How to quantiate extracted DNA before sending it for sequencing?

[jerrybug109]

Hi all,

We just extracted DNA on several samples using Epicenter Masterpure Gram Positive DNA extraction kit. We're going to send it to an NGS service to be library prepped and DNA sequenced, and the NGS service is requesting us perform some quality check on our DNA samples before shipping it to them.

Their criteria are:

DNA sample amount: at least 1.6 micrograms
DNA concentration: at least 20 nanograms/microliter
O.D. 260/280: between 1.8-2.0

It seems that Qubit or Nanodrop allows you to determine DNA concentration and O.D. Are there tools that give you determine the sample amount (in micrograms), or do you just calculate the sample amount from your concentration using simple math?

I'm new to this process - thanks!

- jerrybug109

[nucacidhunter]

Qubit is the best for quantitation for this purpose and total amount is just the simple math. NanoDrop is good for checking potential contaminants left from extraction process and it is normally accurate if concentration is above 15 ng/ul.

[jerrybug109]

Qubit is the best for quantitation for this purpose and total amount is just the simple math. NanoDrop is good for checking potential contaminants left from extraction process and it is normally accurate if concentration is above 15 ng/ul.

Thanks, we have Qubit and both Nanodrop. I'll check it out!

[fpj]

Qubit is best for checking DNA quality. It binds to double stranded DNA and so gives a better indication of yield compared to nanodrop. Nanodrop will always say the yield is higher than Qubit. A lot of sequencing places use qubit as part of their QC process and so it's best to use that than nanodrop.

[kerplunk412]

Qubit is best for checking DNA quality. It binds to double stranded DNA and so gives a better indication of yield compared to nanodrop. Nanodrop will always say the yield is higher than Qubit. A lot of sequencing places use qubit as part of their QC process and so it's best to use that than nanodrop.

I believe you mean DNA quantity. Sorry to be nit-picky but the difference is important here. Nanodrop is actually better for quality because impurities can be seen.

[fpj]

I believe you mean DNA quantity. Sorry to be nit-picky but the difference is important here. Nanodrop is actually better for quality because impurities can be seen.

Oh yes. My mistake!