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Old 08-09-2012, 12:16 PM   #1
Location: East Bay

Join Date: Jul 2012
Posts: 26
Default Yeast gDNA on MiSeq

We are getting only 0.3-0.5 the coverage possible from our MiSeq when sequencing yeast genomic DNA using the Nextera kit for library generation. It seems to be due to tagmentation giving a larger than desired size distribution of fragments. Is yeast DNA a poorer Tn5 target than other DNA? We are using 50 ng DNA (picogreen assay) in the tagmentation reaction, as specified by the protocol. Would reducing the amount of DNA help?
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