Hi,
I have a question about PCR step in library prep. We are using NEBNext kit and we want to pool several samples using index.
After size selection, I have about 250 ng DNA (500 bp including adaptor). It's enough in amount but we need to add index so we still need to do PCR.
The problem is if I only get 4 folds of DNA after PCR, does that means 25% DNA do not get indexed?
Will it affect sequencing efficiency or depth?
Could anyone give me some clue? Thanks a lot.
This is the kit I'm using:
I have a question about PCR step in library prep. We are using NEBNext kit and we want to pool several samples using index.
After size selection, I have about 250 ng DNA (500 bp including adaptor). It's enough in amount but we need to add index so we still need to do PCR.
The problem is if I only get 4 folds of DNA after PCR, does that means 25% DNA do not get indexed?
Will it affect sequencing efficiency or depth?
Could anyone give me some clue? Thanks a lot.
This is the kit I'm using:
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