How many PCR reactions when having different yields of gDNA?

Recently when I extracted the gDNA for PCR, I got different yields between replicates, even though I used the same number of cells for the prep. The difference was up to 50 ug, so if I use all gDNA for PCR, one replicate would have 25 reactions (2.5 ug/reaction) more than the other.

Considering this situation, should I keep the same amount of gDNA for PCR between replicates? Or use all gDNA for PCR but have different numbers of PCR reactions?


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Opal Impabout 2 years ago

I've got a similar question to yours. Do you have the answer so far?

Lemon Mummyabout 2 years ago

The idea is to PCR all the gDNA: so some samples will have more reactions. This is in order to not lose any coverage in your samples.

Magenta Ghoulabout 2 years ago

How many cells do you feed to DNA extraction? Do you remove RNA when extracting gDNA?

Opal Impalmost 2 years ago

I have 50 million cells per sample and I did not remove RNA when extracting gDNA. So would it be better to use all my gDNA products?

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