- Mar 2023
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drive.google.com drive.google.comview1
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testimony
A solemn declaration usually made orally by a witness under oath in response to interrogation by a lawyer or authorized public official. 證據、證明
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- Feb 2023
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socialsci.libretexts.org socialsci.libretexts.org
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No major Articles were added or deleted. The individual property rights of the slave-owner was prioritized. No law could be passed about emancipation. Individuals could not free their own slaves. Immigration of slaves from other states could not be limited. Enslaved people continued to have the right to jury trials, punished as they would be against a White victim, “except in case of insurrection by such slave.”28 But to this was added: “or rape on a white female.”29 The nature of the changes make clear that the principle concern of the members of the secession Convention was to ensure the preservation of slavery.
1861 Slavery differences added to the constitution.
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- Jan 2023
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This sort of policy matches closely to the model page zettelkasten.de which has also a strong focus on memorizing information and excludes secondary elements like vegan food and doing sport for no reason.This is factually incorrect.
reply to u/FastSascha at https://www.reddit.com/r/Zettelkasten/comments/10nolg3/comment/j6naobz/?utm_source=reddit&utm_medium=web2x&context=3
Let those who have not folded an index card to use it as a fork for eating food (vegan or otherwise), throw the first pack of index cards.
Is this the correct zettelkasten translation of John 8:7? Should I number this ZKII, 9/8k?🗃️😉
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www.allocine.fr www.allocine.frVortex1
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Local file Local file
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About twenty thousand of those cards are 3 × 5 inches and seven thousand 5 × 8 inches.
Goitein's zettelkasten is comprised of about 20,000 3 x 5" index cards and 7,000 5 x 8" index cards.
Link to: https://hypothes.is/a/TEiQ5H1rEe2_Amfzi4XXmg
While not directly confirmed (yet), due to the seeming correspondence of the number of cards and their corpus descriptions, it's likely that the 20,000 3 x 5" cards were his notes covering individual topics while the 7,000 5 x 8" cards were his notes and descriptions of a single fragment from the Cairo Geniza.
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- Dec 2022
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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RRID:ZFIN_ZDB-ALT-170801-8
DOI: 10.3389/fphys.2022.1056369
Resource: RRID:ZFIN_ZDB-ALT-170801-8
Curator: @scibot
SciCrunch record: RRID:ZFIN_ZDB-ALT-170801-8
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www.zhihu.com www.zhihu.com
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java中GBK编码格式转成UTF8,用一段方法实现怎么做?
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www.zhihu.com www.zhihu.com
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Unicode 和 UTF-8 有什么区别?
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URL
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soundcloud.com soundcloud.com
- Nov 2022
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www.allocine.fr www.allocine.frL'Avocat1
- Oct 2022
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www.allocine.fr www.allocine.fr
- Sep 2022
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www.reddit.com www.reddit.com
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Posted byu/jackbaty4 hours agoCard sizes .t3_xib133._2FCtq-QzlfuN-SwVMUZMM3 { --postTitle-VisitedLinkColor: #9b9b9b; --postTitleLink-VisitedLinkColor: #9b9b9b; --postBodyLink-VisitedLinkColor: #989898; } I've been on-again/off-again with paper for PKM, but one thing remains consistent each time: I don't enjoy using 4x6 index cards. I much prefer 3x5-inch cards. I realize that it's irrational, but there it is.My question is if I dive into building an antinet, will I regret using 3x5 cards? I already have hundreds of them. I have dividers, holders, and storage boxes for them. I just prefer how they _feel_, as weird as that sounds.I'd like to hear if people are using 3x5 cards successfully or if you've come to regret it.
While it may be slightly more difficult to find larger metal/wood cases for the 4x6 or 5x8 cards, it's a minor nuisance and anyone who wants them will eventually find the right thing for them. Beyond this, choose the card size that feels right to you.
If you don't have an idea of what you need or like, try things out for 10-20 cards and see how it works for you, your handwriting size, and general needs. People have been using 3x5, 4x6, and even larger for hundreds of years without complaining about any major issues. If Carl Linnaeus managed to be okay with 3x5, which he hand cut by the way, I suspect you'll manage too.
Of course I won't mention to the Americans the cleverness of the A6, A5, A4 paper standards which allows you to fold the larger sizes in half to get the exact next smaller size down. Then you might get the benefit of the smaller size as well as the larger which could be folded into your collection of smaller cards, you just have to watch out for accidentally wrapping ("taco-ing") a smaller card inside of a larger one and losing it. I suppose you could hand cut your own 5" x 6" larger cards to do this if you found that you occasionally needed them.
For the pocketbook conscious, 3x5 does have the benefit of lower cost as well as many more options and flexibility than larger sizes.
At least commercial card sizes are now largely standardized, so you don't have deal with changing sizes the way Roland Barthes did over his lifetime.
My personal experience and a long history of so many manuals on the topic saying "cards of the same size" indicates that you assuredly won't have fun mixing different sized slips together. I personally use 3x5" cards in a waste book sense, but my main/permanent collection is in 4x6" format. Sometimes I think I should have done 3 x 5, but it's more like jealousy than regret, particularly when it comes to the potential of a restored fine furniture card catalog. But then again...
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- Aug 2022
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- Jul 2022
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docdrop.org docdrop.org
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when we die we go through eight stages according to the buddhist understanding and each of those stages the first four the elements the sort of solidity if you will i we know they're 01:16:07 not solid but from a conventional perspective the solidity elements the liquidity elements the thermodynamic elements the movement the kinetic elements those all dissolve as we die in 01:16:19 the first four and when that fourth one happens there's no more circulation of blood or of air so we don't breathe we have no circulatory you know blood pressure so we're declared clinically dead but 01:16:30 there's four more stages we go through and those are when the mind becomes successively subtler and those are when we get into the non-dual minds that are the most subtle minds and the last 01:16:43 eighth stage it's called worser in tibetan and we translate that as luminosity or clear light it's not light it's not you know but it's the most utter clear clear mind 01:16:57 and that mind if it goes on if we don't die if we meditate on that luminosity and sustain it through our meditation infinitely we can become a buddha and that's why the buddha is 01:17:09 sometimes called a buddhism an enlightened buddha is a deathless state because you don't actually die so those would be the non-conceptual and non-dual minds and just for completeness 01:17:23 those last four minds are called these are technical terms so it won't make much it won't have much give you much understanding white appearance red increase black near attainment and then this worst air this 01:17:35 luminosity so that's kind of the the the road map if you will for for mine and it's not the brain now on the gross level of thinking in our sensory minds there's a very close 01:17:48 connection with you know meant with the brain okay but when you die the brain is supposed to be dead and you're still alive okay and so these more subtle minds 01:18:01 are not related actually to the brain so we could really say that mind is experience it's awareness it's knowing not knowing something but 01:18:12 the act of knowing so the qualities of mind the most important qualities are awareness and clarity so that gives you just some rough idea of the buddhist understanding of mind or consciousness
Barry gives an explanation of the different levels of mind as the body undergoes death, and particularly, the last 4 of 8 progressively subtler states of mind that are nondual, and therefore, not considered as part of the brain.
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- Jun 2022
- May 2022
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stackoverflow.com stackoverflow.com
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[^[:print:]] will probably suffice for you.**
- FOR ME
- because [ascii] doesnt work in cygwin's grep
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www.justinweiss.com www.justinweiss.com
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You can use a heuristic: only change strings that have one of the bad characters in them, like â. This works well if a character like â won’t ever appear in a valid string. The last time I fixed this kind of bug, though, I wanted to play it safe. I used another useful tool to help: my eyes. Whenever I found a badly encoded string, I printed it out, along with its replacement:
- no magic solutions!
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It seems like those three bytes should be read as UTF-8, where they’d represent a curly quote. Instead, each byte is showing up as a different character. So, which encoding would represent [226, 128, 153] as ’? If you look at a few tables of popular encodings, you’ll see it’s Windows-1252.
-In UTF8 are 3 bytes - In W1252 a byte= a char
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stackoverflow.com stackoverflow.com
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This only forces the client which encoding to use to interpret and display the characters. But the actual problem is that you're already sending ’ (encoded in UTF-8) to the client instead of ’. The client is correctly displaying ’ using the UTF-8 encoding. If the client was misinstructed to use, for example ISO-8859-1, you would likely have seen ââ¬â¢ instead.
- HERE IT IS!
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This answer is not useful Show activity on this post. So what's the problem, It's a ’ (RIGHT SINGLE QUOTATION MARK - U+2019) character which is being decoded as CP-1252 instead of UTF-8. If you check the encodings table, then you see that this character is in UTF-8 composed of bytes 0xE2, 0x80 and 0x99. If you check the CP-1252 code page layout, then you'll see that each of those bytes stand for the individual characters â, € and ™.
- HERE IT IS!
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utf8-chartable.de utf8-chartable.de
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One Latin-1 char per byte
- activar para ver secuencia 2 bytes
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- sequences 2 bytes for no-ascii
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URL
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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DICER1 syndrome encompasses a variety of benign and malignant manifestations including multinodular goitre
Gene: DICER1 PMCID: PMC8451242 PMID: 34552563 Pathogenic Inheritance Pattern: Autosomal Dominant MultipleDiseaseEntities Disease Entity: DICER1 syndrome, multinodular goitre, cystic nephroma, anaplastic renal sarcoma, Wilms tumour, differentiated thyroid carcinoma, gynandroblastoma, ciliary body medulloepithelioma, embryonal rhabdomyosarcoma, pineoblastoma, pituitary blastoma, kidney cyst, pulmonary cyst, Sertoli-Leydig Cell Tumor. Mutation: Germline MultipleGeneVariants Variant & Clinvar IDs: c.3452_3453del (485534), c.316del (no ClinVar ID), c.171_172insAC (no ClinVar ID), c.3434del (no ClinVar ID), c.988C>T (933007), c.5388dup (no ClinVar ID) Zygosity: None provided. Case: At time of operation, the goitre patients living in Denmark were ages 21, 12, 21, 8, 14, and 16. Four underwent total thyroidectomies, and two underwent partial thyroidectomies. The patient originally aged 21 previously had a kidney cyst at age 14 and a pulmonary cyst at an unknown age. The patient aged 14 at time of partial thyroidectomy later manifested a Sertoli-Leydig Cell Tumor at age 15. All six patients were female. CasePresentingHPO: None provided. CasePreviousTesting: thyroidectomy gnomAD: ENSG00000100697.10, https://gnomad.broadinstitute.org/gene/ENSG00000100697 Mutation Type: Frameshift, Nonsense
Tags
- DICER1
- goiter
- c.3452_3453del
- Frameshift
- Gene:DICER1
- c.988C>T
- Cohort
- thyroidectomy
- cancer
- PMID:34552563
- Age:14
- female
- AutosomalDominant
- c.3434del
- goitre
- PMCID:PMC8451242
- MultipleGeneVariants
- Age:8
- Nonsense
- c.5388dup
- Germline
- Age:21
- gnomAD:ENSG00000100697.10
- ClinvarID:485534
- ClinvarID:933007
- c.171_172insAC
- ClinvarID:NotProvided
- Mutation:Frameshift
- c.316del
- Age:12
- Mutation:Nonsense
- Pathogenic
- MultipleDiseaseEntities
Annotators
URL
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stackoverflow.com stackoverflow.com
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This works for me: PowerShell -Command "TREE /F | Out-File output.txt -Encoding utf8"
- WITH POWERSHELL
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You should add this command chcp 65001 before dir command to change code page to UTF-8 @echo off CHCP 65001>nul dir>1.txt Further reading about CHCP command
- DIR NAMES IN UTF-8
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docs.actian.com docs.actian.com
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- hex: 93 y 94
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URL
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www.cl.cam.ac.uk www.cl.cam.ac.uk
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Most European keyboards have keycap labels for the apostrophe and both accents. These have always looked like in the ISO and Unicode standards. The photo below shows the relevant keys highlighted on a standard German PC keyboard, which has the acute/grave accent key left and the number-sign/apostrophe key below the backspace key:
- unicode!
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- Mar 2022
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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RRID:ZFIN_ZDB-ALT-000412-8
DOI: 10.7554/eLife.74821
Resource: (ZFIN Cat# ZDB-ALT-000412-8,RRID:ZFIN_ZDB-ALT-000412-8)
Curator: @scibot
SciCrunch record: RRID:ZFIN_ZDB-ALT-000412-8
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- Dec 2021
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mmcr.education mmcr.education
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spoken or written word.
AND non verbal conduct. Not sure if this counts as a flaw. Its missing a part.
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the Court could also decide that the state doesn't have a legitimate interest in the question of flag desecration
I believe this is incorrect: I think this is the correct reasoning regarding legitimate interest in the question of flag desecration "we do not doubt that the government has a legitimate interest in making efforts to "preserv[e] the national flag as an unalloyed symbol of our country."
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- Nov 2021
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mmcr.education mmcr.education
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dissenting
Concurring
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mmcr.education mmcr.education
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that government may always prohibit the expression of an idea whenever society finds the idea itself offensive or disagreeable.
incorrect, the government may not always prohibit the expression of an idea when...
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mmcr.education mmcr.education
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Simmons-Harris sued, charging that the voucher program violated the First Amendment's free exercise clause because only 10 percent of the private schools available were religious, and only 5 percent of students used their vouchers at private schools.
This area is incorrect, the books version states, "Although no public schools from adjacent districts opted to participate in the program, fifty-six private schools, 80 percent of them religious, did. Religious schools were the choice of the parents of 97 percent of the students who used tuition vouchers to attend private schools."
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mmcr.education mmcr.education
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dissenting
Concurring
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mmcr.education mmcr.education
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offend the 14th Amendment to the Constitution
Incorrect - "Does the voucher program offend the establishment clause..." is the correct legal issue here
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- Oct 2021
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mmcr.education mmcr.education
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dissenting
concurring
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mmcr.education mmcr.education
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Roberts:
incorrect: not a justice on this panel/ case/ trial
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mmcr.education mmcr.education
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Roberts: dissenting
This one didn't happen.
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- Sep 2021
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www.sciencedirect.com www.sciencedirect.com
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ZFIN: ZDB-ALT-020918-8
DOI: 10.1016/j.cub.2021.08.049
Resource: (ZFIN Cat# ZDB-ALT-020918-8,RRID:ZFIN_ZDB-ALT-020918-8)
Curator: @Naa003
SciCrunch record: RRID:ZFIN_ZDB-ALT-020918-8
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s3.us-central-1.wasabisys.com s3.us-central-1.wasabisys.com
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paradigmatic
typical answer to something
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elucidatory
give a clarifying expression
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elucidation.
another word for clarification
Tags
- https://www.google.com/search?q=elucidative&rlz=1C1CHZN_enUS966US966&oq=elucidative&aqs=chrome..69i57j0i512l5j0i10i512j0i512j0i10i30l2.8526j0j4&sourceid=chrome&ie=UTF-8
- https://www.google.com/search?q=elucidation&rlz=1C1CHZN_enUS966US966&oq=elucidation&aqs=chrome.0.69i59i433i512j0i131i433i512j0i433i512j0i512l5j0i10i512j0i512.2598j0j7&sourceid=chrome&ie=UTF-8
- https://www.google.com/search?q=paradigmatic+definition&rlz=1C1CHZN_enUS966US966&oq=paradigmatic&aqs=chrome.1.69i59j0i512l9.4463j1j9&sourceid=chrome&ie=UTF-8
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www.education.gouv.fr www.education.gouv.fr
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les directeurs d'école et les chefs d'établissement doivent communiquer aux associations de parents d'élèves qui en font la demande la liste des parents d'élèves de l'école ou de l'établissement scolaire mentionnant leurs noms et adresses postale et électronique
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- Aug 2021
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www.sciencedirect.com www.sciencedirect.com
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ZDB-ALT-041129-8
DOI: 10.1016/j.devcel.2021.07.021
Resource: (ZFIN Cat# ZDB-ALT-041129-8,RRID:ZFIN_ZDB-ALT-041129-8)
Curator: @Naa003
SciCrunch record: RRID:ZFIN_ZDB-ALT-041129-8
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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RRID:ZFIN_ZDB-ALT-980621-8
DOI: 10.7554/eLife.66078
Resource: (ZFIN Cat# ZDB-ALT-980621-8,RRID:ZFIN_ZDB-ALT-980621-8)
Curator: @scibot
SciCrunch record: RRID:ZFIN_ZDB-ALT-980621-8
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- Jul 2021
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www.sciencedirect.com www.sciencedirect.com
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Zebrafish: Tg(olig2:egfp)vu12
DOI: 10.1016/j.celrep.2019.03.013
Resource: (ZFIN Cat# ZDB-ALT-041129-8,RRID:ZFIN_ZDB-ALT-041129-8)
Curator: @ethanbadger
SciCrunch record: RRID:ZFIN_ZDB-ALT-041129-8
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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Tg(olig2:EGFP)vu12
DOI: 10.1242/dev.105718
Resource: (ZFIN Cat# ZDB-ALT-041129-8,RRID:ZFIN_ZDB-ALT-041129-8)
Curator: @mpairish
SciCrunch record: RRID:ZFIN_ZDB-ALT-041129-8
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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Tg(fli1a:EGFP)y1
DOI: 10.1093/glycob/cwv009
Resource: (ZFIN Cat# ZDB-ALT-011017-8,RRID:ZFIN_ZDB-ALT-011017-8)
Curator: @mpairish
SciCrunch record: RRID:ZFIN_ZDB-ALT-011017-8
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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Tg(fli1:EGFP)y1
DOI: 10.1038/srep11149
Resource: (ZFIN Cat# ZDB-ALT-011017-8,RRID:ZFIN_ZDB-ALT-011017-8)
Curator: @mpairish
SciCrunch record: RRID:ZFIN_ZDB-ALT-011017-8
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www.sciencedirect.com www.sciencedirect.com
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RRID:ZDB-ALT-170927-8
DOI: 10.1016/j.cub.2017.11.018
Resource: (ZFIN Cat# ZDB-ALT-170927-8,RRID:ZFIN_ZDB-ALT-170927-8)
Curator: @evieth
SciCrunch record: RRID:ZFIN_ZDB-ALT-170927-8
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RRID:ZDB-ALT-171002-3
DOI: 10.1016/j.cub.2017.11.018
Resource: (ZFIN Cat# ZDB-ALT-170927-8,RRID:ZFIN_ZDB-ALT-170927-8)
Curator: @evieth
SciCrunch record: RRID:ZFIN_ZDB-ALT-170927-8
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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RRID:ZDB-ALT-130123-8
DOI: 10.7554/eLife.37001
Resource: (ZFIN Cat# ZDB-ALT-130123-8,RRID:ZFIN_ZDB-ALT-130123-8)
Curator: @evieth
SciCrunch record: RRID:ZFIN_ZDB-ALT-130123-8
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www.sciencedirect.com www.sciencedirect.com
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RRID:ZDB-ALT-011017-8
DOI: 10.1016/j.devcel.2017.12.022
Resource: (ZFIN Cat# ZDB-ALT-011017-8,RRID:ZFIN_ZDB-ALT-011017-8)
Curator: @Jmenke
SciCrunch record: RRID:ZFIN_ZDB-ALT-011017-8
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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ZFIN ID: ZDB-ALT-011017-8
DOI: 10.7554/eLife.66596
Resource: (ZFIN Cat# ZDB-ALT-011017-8,RRID:ZFIN_ZDB-ALT-011017-8)
Curator: @evieth
SciCrunch record: RRID:ZFIN_ZDB-ALT-011017-8
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- Jun 2021
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outlier-org.admin.datocms.com outlier-org.admin.datocms.com
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Government Action: Education
First paragraph is Giacomo adaptation, second paragraph is OS text
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www.interviewbit.com www.interviewbit.com
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Angular is nothing but an open-source web application framework. It is being led by the Angular Team at Google and also by a community of individuals and corporations. It is based on TypeScript.Angular is a complete rewrite from the same team that had built AngularJS. It can be used as the frontend of the MEAN stack, which consists of the MongoDB database, Express.js web application server framework, Angular itself (or AngularJS), and the Node.js server runtime environment.
Top Angular 8 Interview Questions to get you ready
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www.sciencedirect.com www.sciencedirect.com
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ZFIN: ZDB-ALT-160119-8
DOI: 10.1016/j.cub.2021.05.042
Resource: (ZFIN Cat# ZDB-ALT-160119-8,RRID:ZFIN_ZDB-ALT-160119-8)
Curator: @Naa003
SciCrunch record: RRID:ZFIN_ZDB-ALT-160119-8
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elifesciences.org elifesciences.org
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ZDB-ALT-011017–8
DOI: 10.7554/eLife.62155
Resource: (ZFIN Cat# ZDB-ALT-011017-8,RRID:ZFIN_ZDB-ALT-011017-8)
Curator: @Naa003
SciCrunch record: RRID:ZFIN_ZDB-ALT-011017-8
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elifesciences.org elifesciences.org
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RRID:ZFIN_ZDB-ALT-011017-8
DOI: 10.7554/eLife.62196
Resource: (ZFIN Cat# ZDB-ALT-011017-8,RRID:ZFIN_ZDB-ALT-011017-8)
Curator: @scibot
SciCrunch record: RRID:ZFIN_ZDB-ALT-011017-8
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Annotators
URL
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- May 2021
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www.sciencedirect.com www.sciencedirect.com
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ZFIN ID: ZDB-ALT-011017-8
DOI: 10.1016/j.cell.2021.04.029
Resource: (ZFIN Cat# ZDB-ALT-011017-8,RRID:ZFIN_ZDB-ALT-011017-8)
Curator: @Naa003
SciCrunch record: RRID:ZFIN_ZDB-ALT-011017-8
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- Mar 2021
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).
AssayResult: 7.3
AssayResultAssertion: Normal
StandardErrorMean: 0.08
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A total of 84 PALB2 patient-derived missense variants reported in ClinVar, COSMIC, and the PALB2 LOVD database were selected
HGVS: NM_024675.3:c.1145G>T p.(Ser382Ile)
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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SUPPLEMENTARY DATA
AssayResult: 58
AssayResultAssertion: Indeterminate
PValue: < 0.0001
Approximation: Exact assay result value not reported; value estimated from Figure 6C.
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SUPPLEMENTARY DATA
AssayResult: -32
AssayResultAssertion: Abnormal
PValue: < 0.0001
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SUPPLEMENTARY DATA
AssayResult: 76.45
AssayResultAssertion: Indeterminate
PValue: 0.0001
Comment: Exact values reported in Table S3.
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To this end, 44 missense variants found in breast cancer patients were identified in the ClinVar database (https://www.ncbi.nlm.nih.gov/clinvar) and/or selected by literature curation based on their frequency of description or amino acid substitution position in the protein (Supplemental Table S1).
HGVS: NM_024675.3:c.23C>T p.(Pro8Leu)
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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Source Data
AssayResult: 83.96
AssayResultAssertion: Not reported
ReplicateCount: 2
StandardErrorMean: 9.89
Comment: Exact values reported in “Source Data” file.
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Source Data
AssayResult: 66.19
AssayResultAssertion: Not reported
ReplicateCount: 2
StandardDeviation: 21.26
StandardErrorMean: 15.03
Comment: Exact values reported in “Source Data” file.
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We, therefore, analyzed the effect of 48 PALB2 VUS (Fig. 2a, blue) and one synthetic missense variant (p.A1025R) (Fig. 2a, purple)29 on PALB2 function in HR.
HGVS: NM_024675.3:c.1250C>A p.(S417Y)
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www.cell.com www.cell.com
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Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function
AssayResult: 1.2
AssayResultAssertion: Abnormal
ReplicateCount: 11
StandardErrorMean: 0.7
Comment: This variant had loss of function of peak current (<10% of wildtype), therefore it was considered abnormal (in vitro features consistent with Brugada Syndrome Type 1). (Personal communication: A. Glazer)
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we selected 73 previously unstudied variants: 63 suspected Brugada syndrome variants and 10 suspected benign variants
HGVS: NM_198056.2:c.1156G>A p.(Gly386Arg)
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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RRID:ZFIN_ZDB-ALT-120221-8
DOI: 10.7554/eLife.64267
Resource: (ZFIN Cat# ZDB-ALT-120221-8,RRID:ZFIN_ZDB-ALT-120221-8)
Curator: @scibot
SciCrunch record: RRID:ZFIN_ZDB-ALT-120221-8
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RRID:ZFIN_ZDB-ALT-041129-8
DOI: 10.7554/eLife.64267
Resource: (ZFIN Cat# ZDB-ALT-041129-8,RRID:ZFIN_ZDB-ALT-041129-8)
Curator: @scibot
SciCrunch record: RRID:ZFIN_ZDB-ALT-041129-8
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elifesciences.org elifesciences.org
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RRID:ZFINID:ZDB-ALT-011017-8
DOI: 10.7554/eLife.63595
Resource: (ZFIN Cat# ZDB-ALT-011017-8,RRID:ZFIN_ZDB-ALT-011017-8)
Curator: @evieth
SciCrunch record: RRID:ZFIN_ZDB-ALT-011017-8
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- Feb 2021
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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RRID:ZFIN_ZDB-ALT-011017-8
DOI: 10.7554/eLife.55792
Resource: (ZFIN Cat# ZDB-ALT-011017-8,RRID:ZFIN_ZDB-ALT-011017-8)
Curator: @scibot
SciCrunch record: RRID:ZFIN_ZDB-ALT-011017-8
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elifesciences.org elifesciences.org
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RRID:ZDB-ALT-130123-8
DOI: 10.7554/eLife.37001
Resource: (ZFIN Cat# ZDB-ALT-130123-8,RRID:ZFIN_ZDB-ALT-130123-8)
Curator: @scibot
SciCrunch record: RRID:ZFIN_ZDB-ALT-130123-8
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URL
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elifesciences.org elifesciences.org
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RRID:ZFIN_ZDB-ALT-011017-8
DOI: 10.7554/eLife.42881
Resource: (ZFIN Cat# ZDB-ALT-011017-8,RRID:ZFIN_ZDB-ALT-011017-8)
Curator: @scibot
SciCrunch record: RRID:ZFIN_ZDB-ALT-011017-8
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jmg.bmj.com jmg.bmj.com
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Supplemental material
AssayResult: 86
AssayResultAssertion: Normal
Comment: See Table S3 for details
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Supplemental material
AssayResult: 6.1
AssayResultAssertion: Abnormal
Comment: See Table S3 for details
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We analysed a total of 82 blood samples derived from 77 individuals (online supplemental table 3). These 77 individuals corresponded either to new index cases suspected to harbour a pathogenic TP53 variant or to relatives of index cases harbouring TP53 variants.
HGVS: NM_000546.5:c.215C>A p.(Pro72His)
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www.bibliaonline.com.br www.bibliaonline.com.br
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visão
No original essa é a palavra marêh
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URL
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10.11.66.200 10.11.66.200
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Last 4 of SSN
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do not put the caller on hold
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- Jan 2021
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journals.biologists.com journals.biologists.com
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ZIRC Cat# ZL1085 ZDB-ALT-011017-8
DOI: 10.1242/dev.193409
Resource: (ZFIN Cat# ZDB-ALT-011017-8,RRID:ZFIN_ZDB-ALT-011017-8)
Curator: @Naa003
SciCrunch record: RRID:ZFIN_ZDB-ALT-011017-8
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- Nov 2020
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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RRID:ZFIN_ZDB-GENO-200128-8
DOI: 10.7554/eLife.60445
Resource: (ZFIN Cat# ZDB-GENO-200128-8,RRID:ZFIN_ZDB-GENO-200128-8)
Curator: @evieth
SciCrunch record: RRID:ZFIN_ZDB-GENO-200128-8
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- Sep 2020
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ecourse.auca.kg ecourse.auca.kgFYS.pdf3
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of which the handle [was x minas] of gold, he provided for his friend.
It shows the his friend Enkidu.
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o Enkidu, may the paths [of] the Forest of Cedar mourn you [without pause,] by day and by night!
Message of Gilgamesh to Enkidu when he died.
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Gilgamesh [began mourning] his friend:
It shows their friendship.
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www.education.gouv.fr www.education.gouv.fr
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À cette fin, comme le prévoit l'article D. 111-8 du Code de l'éducation, les directeurs d'école et les chefs d'établissement doivent communiquer aux associations de parents d'élèves qui en font la demande la liste des parents d'élèves de l'école ou de l'établissement scolaire mentionnant leurs noms, adresses postale et électronique, à la condition que ceux-ci aient donné leur accord exprès à cette communication.
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- Jan 2020
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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RRID:ZFIN_ZDB-ALT-020918-8
DOI: 10.7554/eLife.46275
Resource: (ZFIN Cat# ZDB-ALT-020918-8,RRID:ZFIN_ZDB-ALT-020918-8)
Curator: @evieth
SciCrunch record: RRID:ZFIN_ZDB-ALT-020918-8
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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RRID:ZFIN_ZDB-ALT-011017-8
DOI: 10.7554/eLife.42881
Resource: (ZFIN Cat# ZDB-ALT-011017-8,RRID:ZFIN_ZDB-ALT-011017-8)
Curator: @evieth
SciCrunch record: RRID:ZFIN_ZDB-ALT-011017-8
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reg.clinicalgenome.org reg.clinicalgenome.org
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allele id lookup result: http://reg.clinicalgenome.org/redmine/projects/registry/genboree_registry/by_caid?caid=CA398917343
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www.sciencedirect.com www.sciencedirect.com
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8
Monarch lookup result: https://monarchinitiative.org/phenotype/HP:0001763
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8
Monarch lookup result: https://monarchinitiative.org/phenotype/HP:0001382
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8
Monarch lookup result: https://monarchinitiative.org/phenotype/HP:0002705
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8
Monarch lookup result: https://monarchinitiative.org/phenotype/HP:0000348
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muscular hypotonia
Monarch lookup result: https://monarchinitiative.org/phenotype/HP:0001252
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motor development
Monarch lookup result: https://monarchinitiative.org/phenotype/HP:0001270
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speech and language development
Monarch lookup result: https://monarchinitiative.org/phenotype/HP:0000750
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- Jul 2019
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www.milestonedocuments.com www.milestonedocuments.com
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Nobel Peace prize
What did he do to win the noble prize?
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We deplore all forms of violence
They are non-violent
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URL
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www.milestonedocuments.com www.milestonedocuments.com
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last war
Meaninng World War 2
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URL
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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Drugs and chemicals
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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Statisticalanalysis
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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TLC study of alkaloids
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TLC study of phenols
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TLC study of alkaloids
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Activation of TLC plate
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Preparation of TLC plate
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Study of Phytochemicals by Thin Layer Chromatography (Wagner, and Bladt, 1996)
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Test for cardiac glycosides
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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RAPDand SSRscoring and data analysis
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Assayof xylanase activity
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Dinitrosalicylate reagent (DNS)(per liter)
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Citrate phosphate buffer
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Reagents
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Xylanase activity
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Evaluation of bioagents against the pathogen
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Laboratory screening of antagonists against the test pa
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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Production of caseinase
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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Gel documentation and image analysis
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Detection of toxR gene
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Detection of tlhgene
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Extraction of genomic DNA
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Detection of V. parahaemolyticusspecies-specific gene
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Isolation of V. parahaemolyticuson HiCrome Vibrio
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Isolation and identification of V. parahaemolyticus
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Gelatinase production
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Gelatinase productio
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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Distribution of antibiotic resistance genes in Vibriofrom Cochin estuary, shrimp farm and seafood
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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Reaction resuspension
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Post-reaction clean up
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Sequencing PCR
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Sequencing reaction
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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Python Molecular Viewer
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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Quantification of MMP or ΔΨmloss
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Assessmentof mitochondrial membrane potential(MMP or ΔΨm)
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Glutathione peroxidase (GPx) assay
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Superoxide dismutase (SOD) assay
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Catalase assay
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Catalytic activity
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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Estimation of bioaccumulation of trace metals inmuscle and gill tissue of fish, Labeo rohita
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- Jun 2019
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krishikosh.egranth.ac.in krishikosh.egranth.ac.in
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Grain yield per plot (g)
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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Post vaccination enhanced expression of activation marker CD69 and Chemokine receptor CCR5 on memory B cells in HBV positive newborns
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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Freezingor storageof cells
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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Cytoskeleton extraction buffer (CSE)
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Ni-NTA pull down
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RNA Interference
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pET30a-β-catenin-FL (Antigen for -catenin antibody)
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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Protein estimation of sonicate
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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2 Bioassay
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Proliferation Assay
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Target binding assay
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Cytotoxicity assay
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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ACK lysis Buffer
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lgG-isotyping in the sera of the immunized animals
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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Polymerase chain reaction (PCR)
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Genomic DNA extraction
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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RT-PCR analysis
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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Ectomorphic endomorph:
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Femur
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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concentrators (Amicon), and subjected to reduction with 0.0.5 M sodium dithionite. For this an appropriate amount of anaerobically prepared dithionite solution was added to the reconstituted Hb and the reaction mixture was quickly passed through a Sephadex G25 gel filtration column (30 em x 1.5 em) equilibrated with 0.05 M Tris HCI (pH 7.4), in order to minimize the duration of contact of dithionite with the protein. The reduced Hb was dialyzed extensively against 0.01 M potassium phosphate buffer (pH 6.5) and loaded onto a CM52 column (1 Ocm x 1.5cm) equilibrated with the same buffer. A linear gradient of 150 ml each of 0.01 M potassium phosphate buffer (pH 6.5) and 0.015 M potassium phosphate buffer (pH 8.5) was employed to elute the protein from the column
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Construction of mutant a globins
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Reconstitution of a globin and rf chain into HbS tetramers
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- May 2019
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oll.libertyfund.org oll.libertyfund.org
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Paduans
Paduans is a Venetian Italian city that was founded by the Trojan price Antenor in 1185 B.C after the destruction of Troy. Antenor who was the founder of the city was said to be a tradior of the Trojans and he delivered Palladio who was the tailsmans of troy to Odysseus and Diomedes and in exchange he recieved salvation for himself and his family. Today Paduans in known to be one of the most beautiful cities in Veneto.
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URL
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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The reaction mixture contained 0.2 mL of enzyme sample, 0.3 mL of buffer and 0.5 mL of p-nitrophenyl-β-D-glucopyranoside (1.0 mM) prepared in 100 mM buffer as the substrate. The reaction was terminated after 30 min of incubation at 70 °C by adding 2 mL of sodium carbonate-bicarbonate buffer (0.1 M, pH 10.0). The liberation of p-nitrophenol was measured at 400 nm and its yield was determined using a standard curve of p-nitrophenol (1-10 μg mL-1) prepared in sodium carbonate-bicarbonate buffer
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β-Glucosidase
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The activities ofβ-xylosidase, xylan acetylesterase and arbinofuranosidase were measured using 1 mM p-nitrophenylxylopyranoside, p-nitrophenylacetate and p-nitrophenylarabinofuranoside, respectively prepared in sodium citrate buffer (0.1 M, pH 7.0). One mL of reaction mixture containing 0.2 mL of crude enzyme solution, 0.3 mL of sodium citrate buffer (0.1 M, pH 7.0) and 0.5 mL of substrate was incubated at 80 °C for 30 min. The reaction was terminated by adding 2 mL sodium carbonate-bicarbonate buffer (1.0 M, pH 10.0). The activities were determined using p-nitrophenol standard curve (1-10 μg mL-1) drawn using absorbance values measured in spectrophotometer at 400 nm. One unit of the enzyme is defined as the amount of enzyme that liberates 1μmole of p-nitrophenol mL-1min-1 under assay conditions.
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Assays for β-Xylosidase, acetylesterase and arbinofuranosidase
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Xylanolytic activity was determined according to Archana and Satyanarayana (1997). The reaction mixture containing 0.5 mL of 1% birchwood xylan in glycine NaOH buffer (0.1 M, pH 9.0) and 0.5 mL of cell free sonicated supernatant was incubated at 80 °C in a water bath for 10 min. After incubation, 1 mL DNSA reagent (Miller, 1959) was added to the reaction mixture and the tubes were incubated in a boiling water bath for 10 min, followed by the addition of 400 μL of 33% w/v sodium potassium tartrate. The absorbance values were recorded at 540 nm in a spectrophotometer (Shimadzu, Japan). The liberated reducing sugars were determined by comparing the absorbance values of these with a standard curve drawn with different concentrations of xylose. One unit (IU) of xylanase is defined as the amount of enzyme required for liberating one μmol of reducing sugar as xylose mL-1 min-1under the assay conditions. Composition of Dinitrosalicylic acid (DNSA) reagent NaOH - 10.0 g Phenol - 2.0 g DNSA - 2.0 g Distilled Water - 1000 mL DNSA reagent was stored in an amber bottle at 4 °C till further use. Sodium sulphite (0.05 % v/v) was added just before the use of the reagent.
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Enzyme Assays
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A stock solution of xylose (1 mg mL-1) was prepared in distilled water. A dilution series ranging from 100-1000 μg mL-1 was prepared from the stock solution. To 1 mL of solution, 1mL of DNSA was added and kept in a boiling water bath for 10 min and then 400 μL of sodium potassium tartrate solution was added and kept it for cooling. The absorbance was recorded in a spectrophotometer (Shimadzu, UV-VIS) at 540 nm
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The clear cell-free supernatants were used as the source of crude recombinant xylanase.
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Preparation of standard curve of xylose
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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Tris, pH 7.4, 1 mM dithiothreitol, and 10% glycerol. Protein concentration w~s detennined by densitometry analysis of Commassie stained gels. Protein samples were stored at -70°C until further use
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To facilitate the expression of recombinant GST-CDPK4 or its mutants, the desired regions of enzyme were PCR amplified using pGEMT-PfCDPK4 as template and PCR primers which possessed overhangs for XhoI and SmaI restriction enzymes (see List II). Often, the PCR products were cloned in TA cloning vector pGE¥T-I easy. Clones in pGEMT-easy vectors were digested with appropriate restriction enzymes to release the inserts. The released inserts were cloned in expression vector pGEX4T-l to facilitate the expression of recombinant proteins. In some cases, the PCR products were digested directly with restriction enzymes and ligated into expression vectors. The plasmid DNA for expression was used to transform E. coli BL21-RIL (Stratagene) strain for the expression of GST-PfCDPK4 and its mutants. Protein expression was induced by overnight incubation of cells with O.lmM IPTG at 18-20°e. Subsequently, cell pellets were suspended in ice cold lysis buffer, contaiJ;1ing 50 mM Tris, pH 7.4, 2 mM EDTA, 1 mM dithiothreitol, 1% TritonX-100, and protease inhibitors (lmM phenylmethylsulfonyl fluoride, 10~g/ml leupeptin, 1 O~g/ml pepstatin) and sonication was performed for 6 cycles of one minute each. The resulting cell debris was removed by centrifugation at 20,000g for 40 min at 4°C. Fusion proteins from the cell lysates were affinity-purified using glutathione-sepharose resin (Arnersham). Briefly, after the protein binding, the resin was washed with lysis buffer, and bound proteins were eluted with 50 mM Tris, pH 8.0 with 10 mM glutathione. Finally, purified proteins were dialyzed against 50 mM
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xpression and Purification of Recombinant GST (Glutath ion e-S-Transferase) fusion PfCDPK4 and its mutant
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For synchronization, mostly ring stage parasites (10 to 12 h post-invasion) wen~ used. The parasite culture was centrifuged at 200g for 5min and the supernatant was discarded. To the pellet, 4 ml of 5% sorbitol was added, mixed gently and incubated for 15min at 37°C. The mix was shaken 2 or 3 times and centrifuged at 200g followed by washing 3 times in complete medium (list I). The culture was then maintained at 5% hematocrit in a 37°C incubator
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orbitol-synchronization 0/ P./alciparum
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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THP-1 macrophages and human peripheral blood monocyte derived macrophages were transfected with SMARTpool Bcl-2 siRNA (15 pmol), or ER-a siRNA (100 pmol), or ER-~ siRNA (100 pmol), or with negative control siRNA (15 pmol or 100 pmol) using TranspassR2 transfection reagent. Prior to transfection, the cells were depleted of serum by washing 2x with serum-free media. The transfection complex was prepared by diluting 0.5 J!L of transfection reagent A and 1.0 J!L of transfection reagent B to 400 J!L of serum-free media and siRNA's were added to the mix at an appropriate concentration and incubated for 20 min at room temperature. The formed transfection complexes were transferred gently using a large bore pipette tip to 105 cells/well grown in 24 well plates and incubated for 6 h, following which fresh complete medium was added. Transfection efficiency was estimated by observing Cy3-fluorescence of the negative control siRNA with a Nikon TE2000E fluorescence microscope using a tetramethyl rhodamine filter (530-580 nm). For all transfections, target protein knockdown was assessed 24 h after transfection by probing extracts oftransfected cells on Western blots using appropriate antibodies
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siRNA transfection
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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resuspended in 0.2 ml TE, and extracted successively with phenol, phenol-chloroform, and chloroform. In the aqueous phase, 0.25 volume of J 0 M ammonium acetate and two volumes of chilled ethanol were added and the mixture was incubated at room temperature for 5 min. to precipitate the plasmid DNA. The pure plasmid DNA was recovered by centrifugation at J 2,000 g for J 0 min. at 4 °C, washed with 70 %ethanol, dried and resuspended in TE buffer (pH 8.0). The amount and the purity of the DNA was done spectrophotometrically by recording the absorbance at 260 nm.
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Plasmid DNA was prepared by alkaline lysis method of Ish-Horowicz ( 1981 ). 5 ml cultures were grown as described for small scale plasmid preparation. 0.5 ml from the growing culture was inoculated into 250 ml of LB containing ampicillin. The culture was grown for 12 h at 37 °C with vigorous shaking, centrifuged at 3000 g, at 4 °C, for 15 min. and the bacterial pellet was resuspended gently in 1 0 ml TEG buffer. The mixture was incubated at room temperature for 10 min., followed by addition of 20 ml of freshly prepared alkaline-SDS solution. The contents were mixed by inversion and the mixture was kept on ice for 10 min., followed by the addition of 15 ml of chilled potassium acetate solution. The contents were mixed by inverting the tube, and incubated on ice for 10 min. The lysed cell suspension was centrifuged at 5000 g, at 4 °C, for 20 min. The supernatant was taken, and nucleic acids we~,-.. precipitated by adding 0.6 volume of chilled isopropanol. The mixture was incubated on ice for 10 min. followed by centrifugation at 5000 gat 4 °C, for 10 min .. The pellet was washed with 70% ethanol, dried and resuspended in TE buffer. The plasmid DNA was purified further to remove the contaminating proteins and RNA following the PEG purification protocol as described by Sambrook et al ( 1989). Equal volume of chilled 5 M lithium chloride solution was added to DNA . suspension, mixed well and incubated on ice for 10 min. The precipitate was removed by centrifugation at 10,000 g at 4 °C, for 10 min. DNA was precipitated from the supernatant by adding equal volume of isopropanoL The mixture was centrifuged at 10,000 g for 10 min. at 4 °C and the pellet was washed with 70% ethanol. The DNA thus obtained was incubated in TE buffer containing 20 J.tg/ml of DNase free RNase A for 30 min. at room temperature. Afterwards, equal volume of 1.6 M NaCl containing 13% (w/v) PEG 8000 was added to DNA solution. The contents were. thoroughly mixed and centrifuged at 10,000 g, at 4 °C, for 10 min .. The pellet was
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Large Scale Plasmid DNA Preparation
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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administered as and when required. Animals were put on continuous mating with males of proven fertility after administration of the three primary injections and monitored for menstrual cyclicity and conception. Ab titres were determined as described above except that anti-monkey HRPO conjugate was used as the revealing Ab.
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Female bonnet monkeys (Macaca radiata) reared at the Primate Facility (Nil, New Delhi) were selected and serum progesterone levels were estimated for atleast three months in samples which were collected biweekly. Animals showing atleast two consecutive normal ovulatory peaks (serum progesterone levels >2 ng/ml) (Bamezai, 1986) were selected for fertility trials. Five animals (MRA 375, 515, 640, 672, 770) immunized with 250 Jlg equivalent of r-bZP3, expressed in SG I3009[pREP4] cells, conjugated to DT, was emulsified with Squalene and Arlacel A, adjuvants permitted for human use, in a ratio of 4: I and administered intramuscularly at two sites. In addition, the primary dose also contained I mg/animal of SPLPS as an additional adjuvant. Animals were boosted at intervals of 4-6 weeks depending on the Ab titers with 250 Jlg of r-bZP3-DT using Squalene and Arlacel A as adjuvants. A second group of 3 monkeys (MRA 384, 502, 661) were immunized using a slightly different protocol. The primary immunization consisted of 125 J.lg of r-bZP3-DT and 125 J.lg of r-bZP3-TT (expressed in BL2I(DE3) cells) using the same adjuvants and immunization protocols mentioned above except that boosters were administered alternately with 250 Jlg of r-bZP3-DT or -TT conjugates using Squalene and Arlacel A as adjuvants. Following completion of the primary immunization and 2 boosters at monthly intervals, bleeds (1-2 ml) were collected biweekly from the antecubital vein for estimation of progesterone levels and Ab titres. Boosters were
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Immunization of Female Bonnet Monkeys
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Radiolabeling of Recombinant Proteins in Infected S/9 Cells
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Peptide antisera were generated in the laboratory against peptides PI, 23-45 aa residues with an extra lysine at the N-terminus (KQPFWLLQGGASRAETSVQPVL VE), P2, 300-322 aa residues (CSFSKSSNSWFPVEGPADICQCC) and P3, 324-347 aa residues (KGDCGTPSHSRRQPHVVSQWSRSA) corresponding to bZP3 precursor protein in rabbits and were used to determine their reactivity with the r-bZP3 protein expressed in E. coli in an enzyme linked immunosorbent assay (ELISA). Microtitration plates were coated with 200 ng of r-bZP3 or I J.tg/well of the peptide. HRPO conjugated goat anti-rabbit Ig at I :5000 dilution was used as revealing Ab.
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Reactivity with Anti-peptide Sera
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The bZP3 sequence was analyzed using PCgene and Lasergene DNA and protein analysis softwares. The alignment of the bZP3 aa sequence with the homologous sequences from other species was carried out using the Cluster V Multiple Alignment Programme (Higgins and Sharp, 1989).
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Analysis of Sequence
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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Transformation was performed in chilled 1.5 ml eppendorf tubes, using 200 ul of competent cells and about 50 ng of ligated plasmid DNA. Frozen competent cells were thawed in ice and the DNA was added immediately after thawing. The DNA volume was always kept under 30 ul. The DNA was mixed well with the cells by gentle tapping, and the tube incubated in ice for 3 0 minutes with occasional gentle shaking. The tube was then immersed in a 42°C water bath for 2 minutes, to give a heat shock to the cells. The cells were then incubated in ice for 10 minutes. Next, 1 ml LB was to the cells, and the cells incubated in a 37°C water bath without shaking, for one hour. 50 ul aliquots were plated in triplicate from the transformed cell mixture on suitable antibiotic containing agar plates and incubated 0/N at 37°C to select the transformants. In case of JM105 cells, the transformed cells were plated on antibiotic containing agar plates on which 50 ul of 2 % X-gal ( made in dimethyl formamide ) , and 10 ul of 100 mM IPTG had been spread in advance, to select for the lac-phenotype. The lac-colonies appeared colourless while the lac+ colonies were blue. For each batch of transformations, a negative control was included in which no DNA was added to the cells while keeping the rest of the procedure the same as for the test transformations.
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Transformation procedure.
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were stored at -70°C for at least six months without any significant loss in the competence.
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