SEQanswers

SEQanswers (http://seqanswers.com/forums/index.php)
-   RNA Sequencing (http://seqanswers.com/forums/forumdisplay.php?f=26)
-   -   TruSeq Stranded Total RNA + NEB Barcodes?? (http://seqanswers.com/forums/showthread.php?t=76020)

RickC7 05-16-2017 09:19 AM

TruSeq Stranded Total RNA + NEB Barcodes??
 
Looking to get more index possibilities out of a TruSeq Stranded Total RNA LT kit. NEB Ultra directional workflow looks the same and NEB will sell the extra barcode kit alone. As far as I can tell, it looks like the NEB index adapters will work with the TruSeq kit, since both use dA-tailing... NEB #E7335, #E7500. Anyone have experience with this?

Thanks!

MU Core 05-17-2017 05:39 AM

Keep in mind you'll require treatment of the adapter ends with the USER enzyme to free the circularized ends for downstream sequencing chemistry. This is a step you would have to incorporate into the TruSeq workflow. Not sure how easy that would be with the master mixes, etc. of the TruSeq kit.

RickC7 05-17-2017 06:01 AM

Yes, I found a couple of caveats and work-arounds, but thankfully I won't need to employ any customization as Illumina gratefully offered to send me extra barcodes for this instance.
Thanks!


All times are GMT -8. The time now is 07:17 PM.

Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2020, vBulletin Solutions, Inc.