TruSeq Stranded Total RNA + NEB Barcodes??

RickC7 · 05-16-2017, 09:19 AM

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!

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05-16-2017, 09:19 AM	#1
RickC7 Member Location: Baton Rouge, Louisiana Join Date: Feb 2010 Posts: 31	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!

05-17-2017, 05:39 AM	#2
MU Core Member Location: Columbia, Missouri Join Date: Apr 2008 Posts: 57	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.

05-17-2017, 06:01 AM	#3
RickC7 Member Location: Baton Rouge, Louisiana Join Date: Feb 2010 Posts: 31	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!