SEQanswers

SEQanswers (http://seqanswers.com/forums/index.php)
-   Metagenomics (http://seqanswers.com/forums/forumdisplay.php?f=29)
-   -   Concentrations of Post-indexed libraries (http://seqanswers.com/forums/showthread.php?t=72680)

molbionerd 11-21-2016 11:36 AM

Concentrations of Post-indexed libraries
 
Hello all

I have been working with microbiome libraries using Illumina's recommended protocol and sequencing the V3-V4 regions. My question is about the post-indexed library concentrations. I am getting wildly variable concentrations of the indexed samples (measured by qubit, ranging from too low to be detected to 38 ng/uL) with the expected size (~630 bp). I'm just curious what other people typically see for their concentrations at this step. We are following the Illumina protocol and using 8 cycles of their recommended conditions. Should we increase the number of cycles at this step to increase our concentrations? What about increasing the amount of starting template from 5 uL to 10 uL or 15 uL? Any advice is greatly appreciated.


All times are GMT -8. The time now is 03:10 AM.

Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2019, vBulletin Solutions, Inc.