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Old 04-03-2017, 10:29 AM   #1
Amcb1
Junior Member
 
Location: UK

Join Date: Apr 2017
Posts: 4
Default Unexpected 425bp band un Truseq stranded mRNA library prep

Hello,
I joined the community earlier today and I have a problem, I have carried out a Truseq Stranded mRNA HT library prep. and have an unexpected band in some of the samples. I used half of the same kit a few weeks ago with no problems and good sequencing. In other similar preps with the HT and LT kit the odd band has not been present, I have previously, occasionally seen adapter dimers at around 120 bp, which is explainable. This band is around 425 bp and is not present in all of the preps and is not present in the mock (water only) control, which makes me think that it is not in the core reagents. It looks like an amplicon, as it has a very sharp peak on the Tape station, and does amplify when I diluted 1:1000 and carried out a normal PCR with KAPA primers for 10 cycles. I am attaching the Tape station results for all of the libraries. If anyone has seem something similar or has any ideas I would be very grateful. At present my strategy will be to try a couple of samples on a nano cartridge on a Miseq and see if any of the results leap out as adapters or something unexpected.
With hope
A
Attached Files
File Type: pdf 2017-03-30-02.pdf (1.32 MB, 30 views)
File Type: pdf 2017-03-30-01.pdf (1.29 MB, 7 views)
File Type: pdf 2017-03-30-03.pdf (678.9 KB, 5 views)
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