Restriction Digestion
See notes in this section for restriction digestion tips
Restriction Digestion
See notes in this section for restriction digestion tips
DNA fragmentation by restriction digestion prior to dropletgeneration enables optimal accuracy by separating tandemgene copies, reducing sample viscosity, and improving templateaccessibility for input samples >66 ng per well
NEB says
Digestion is recommended whenever DNA input is greater than 75 ng Source
NEB says that biorad recommends these enzymes: AluI, CviQI, HaeIII, HindIII-HF, MseI
More guidelines
- Assemble ddPCR reactions at room temperature, this allows the restriction enzyme to digest the gDNA during the reaction set-up period
- Prepare reaction mixes as you would for a standard ddPCR reaction. Add 0.5–1 μL of each restriction enzyme (5–20 units, depending on enzyme concentration) to the reaction mixture
- After set-up, simply continue droplet generation as normal
- Restriction enzyme will be inactivated during first PCR denaturation step
Refer to this protocol mentioned in the BioRad page, annotation
Mayer says time-restricted eating — a form of intermittent fasting that requires you to squeeze all your daily calories into a compressed feeding window — may be helpful. “The migrating motor complex is rarely mentioned in these articles on intermittent fasting, which is surprising because it’s so well-studied,” he says.To ensure the MMC has enough time to perform its duties, aiming for 14 hours without caloric foods or drinks is a good target, he says. For example, you could avoid all calories between 8 p.m. and 10 a.m. “The 14 hours without food intake would allow the MMC to kick in and not only cleanse your gut of any undigestible, unabsorbable food components, but also to reestablish the normal proximal-to-distal gradient of gut microbial density,” he says.
!- For : microbiome health - fasting for 14 hours helps the migrating motor complex (MMC) maintain gut health
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There are a few things we know that promote longevity and healthspan. Caloric restriction and intermittent fasting are one of the most effective ways of doing so as shown by research in virtually all species.
This could explain why Dr Clemens' [[PKD]] protocol has me around 1,000 calories a day
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The weakest first factor accounted for 18.3% of variance
This factor may be weak because the sample consists of Sudanese gifted children, which may have restricted the range of correlations in the dataset.