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Old 05-22-2014, 11:07 AM   #1
skerker
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Location: CA

Join Date: Nov 2009
Posts: 10
Default Problems with gDNA prep for PacBio

Our goal is to prep genomic DNA from S. cerevisiae to make either a 10kbp or 20kbp library. We have been having trouble with sample quality (either contamination or damage?) resulting in short PacBio reads (only about 1-2kbp). The library "looks good" - it was the expected size ~10kbp - but something else must be wrong...

It seems that very high quality gDNA is needed for good PacBio results. If anyone has a protocol that they would be willing to share for fungi, I'd appreciate it!

We have tried, Qiagen and Zymo research products, in addition to old-school glass beads / phenol-chloroform extraction.

Best,
Jeffrey Skerker
UC Berkeley
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