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Old 06-10-2015, 02:11 PM   #1
yuehuang
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Default Nextera XT library products low average bp size

Dear All,

I'm working on some ancient blood samples(about 20yrs old), trying to do some metagenomic studies on microbiota. The issue is: most of the library prep products' average bp falling around 250 like what was attached here. Is this MiSeqable? Your ideas and suggestions are more than welcome!

Tons of thanks!

Yue

PS: the library was prepared through the Nextera XT kits, and the tagmentation time was 2:30 minutes, amplified by 15 cycles. The others were strictly following Nextera XT protocol.

PPS: DNAs were probably degraded, but I randomly bioanalyzed the DNAs, some of their average bp were truly very low, lower than 250, but we did get some DNAs between 180-1100 bp, nevertheless, the library prep products are the similar as attached.
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Old 06-10-2015, 07:41 PM   #2
nucacidhunter
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It can be sequenced. Because average insert size is short (50-100 bp), I would use 150 cycle v3 kit for sequencing. Average quality would look low because most fragments will sequence through adapter but trimming should improve it.
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Old 06-11-2015, 01:06 PM   #3
yuehuang
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Quote:
Originally Posted by nucacidhunter View Post
It can be sequenced. Because average insert size is short (50-100 bp), I would use 150 cycle v3 kit for sequencing. Average quality would look low because most fragments will sequence through adapter but trimming should improve it.
Thanks much! I will give it a shot, will keep you and this thread posted. Hopefully this try will help others.
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