SEQanswers - View Single Post - Library PCR enrichment questions (with image!)

Heisman · 06-06-2011, 02:14 PM

If the PCR works then the adapter ligation had to work to some extent.

Is this an Illumina library prep? I've routinely used 15-30 minute adapter ligations with no issues.

How much starting material did you use? And what manual did you follow? Were there any modifications?

SYBR gold stains single-stranded DNA a lot better than EtBr, I believe, but otherwise I can't think of a reason why it would run differently.

06-06-2011, 02:14 PM	#4
Heisman Senior Member Location: St. Louis Join Date: Dec 2010 Posts: 535	If the PCR works then the adapter ligation had to work to some extent. Is this an Illumina library prep? I've routinely used 15-30 minute adapter ligations with no issues. How much starting material did you use? And what manual did you follow? Were there any modifications? SYBR gold stains single-stranded DNA a lot better than EtBr, I believe, but otherwise I can't think of a reason why it would run differently.