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Old 08-21-2016, 10:26 AM   #1
Vinn
Member
 
Location: Sweden

Join Date: Nov 2014
Posts: 18
Default Please help! DNA samples for whole genome sequencing

Hi folks,

I have extracted DNA from fungi for whole genome sequencing (Truseq PCR free with 350 bp insert and HiSeq 2500 PE) using a kit. The extraction went smooth until I realized at the end that I treated my samples with only half of the recommended amount for RNase A (2 ul of 10 mg/ml instead of 2 ul of 20 mg/ml). I tried checking the DNA samples on 1% agarose and following is the gel image with 100-1000 bp ladder.





I am not sure there are RNA bands (very faint though) at around 100-200 bp. Do you think my DNA samples are good enough to go for whole genome sequencing without additional treatments? Or shall I try to treat them with RNase A again? Unfortunately, I run out of fresh samples.

I appreciate all your comments and suggestions.

Best
Vinn
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Old 08-21-2016, 10:40 AM   #2
jdk787
josh kinman
 
Location: Austin

Join Date: Apr 2014
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These look fine. Even if you have some RNA in your sample it won't make into your final library after fragmentation and ligation steps.
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dna extraction, rna dna contamination, whole genome sequencing

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