Hi everyone,
Lately I have been extracting DNA to do genotyping-by-sequencing, and one of the requirements of the company that is receiving the samples is that DNA is at a minimum concentration of 50 ng/ul. After a first look at the gel, I was sure I had recovered a lot of DNA, but after quantifying with Qubit (dsDNA, broad range) I get results that are below 50 ng/ul for most of the samples. I tried using different Qubit kits or different incubation times, and the results are very similar. Other colleagues have looked at the gel and the readings and agreed that the results are odd.
I attach an example of part of the samples. Most of them seem good (they have high molecular weight) but the readings are mostly from 20-40 ng/ul.
Most of these samples had very low readings for NanoDrop ratios, indicating probable protein contamination, but I thought that it would not be a problem for the Qubit. Can an excess in contaminants impede the procedure, or do the samples actually have low DNA yield?
Lately I have been extracting DNA to do genotyping-by-sequencing, and one of the requirements of the company that is receiving the samples is that DNA is at a minimum concentration of 50 ng/ul. After a first look at the gel, I was sure I had recovered a lot of DNA, but after quantifying with Qubit (dsDNA, broad range) I get results that are below 50 ng/ul for most of the samples. I tried using different Qubit kits or different incubation times, and the results are very similar. Other colleagues have looked at the gel and the readings and agreed that the results are odd.
I attach an example of part of the samples. Most of them seem good (they have high molecular weight) but the readings are mostly from 20-40 ng/ul.
Most of these samples had very low readings for NanoDrop ratios, indicating probable protein contamination, but I thought that it would not be a problem for the Qubit. Can an excess in contaminants impede the procedure, or do the samples actually have low DNA yield?
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