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Old 09-30-2020, 10:25 AM   #1
Ggnome
Junior Member
 
Location: San Diego

Join Date: Jul 2020
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Question WGS library gen gone wrong?

Hi, I'm doing a WGS sequencing project. The original samples provided were whole human blood in EDTA. After DNA extraction and library gen with the TruSeq DNA PCR-Free protocol w/ 350 bp insert, I got the following results on Tapestation (gel and histogram of one of the 5 libraries, which looks similar to them all):

https://imgur.com/a/T0CJLcS

The result looks a lot different from the figures in the Illumina Library QC whitepaper, so I was wondering if anyone knows if it's a good idea to proceed to sequencing now with this result? If not, any ideas on what went wrong?

I'm concerned because I need to save project budget for the sequencing itself, so I'd rather re-do the DNA extract and library gen than sequence bad libraries and get bad data.

Thanks in advance!
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Old 10-01-2020, 04:29 AM   #2
jdk787
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As long as the qPCR results are good those libraries should be fine to sequence.
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Old 10-01-2020, 10:09 AM   #3
luc
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You might want to have a look at this FAQ:
https://dnatech.genomecenter.ucdavis...ree-libraries/
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Old 10-01-2020, 03:32 PM   #4
Ggnome
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Thanks so much for your replies jdk787 and luc! This UC Davis Q&A is very helpful.
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