Hi there
I've been using the Illumina Truseq small RNA kit, to little success. Starting off with 1ug of total RNA (all with RIN values of 10 I should add!), I took the samples through the whole library prep process, ran them on the polyacrylamide gel (see attached gel pic) and saw faint bands. Unfortunately, when I ran the finished, purified libraries my Bioanalyser results were very substandard (see attached .pdf). Most of them were almost flat lines!! Only one (sample #2) looked vaguely decent (this being the library with the brightest bands on the gel).
I was wondering whether anybody else has had similar problems with this? I have followed the protocol completely, and left the samples to elute for ~18 hours - the protocol says you can leave them in the shaker overnight, so I would expect higher yield if anything!
I also ran the libraries pre-PAGE on the Bioanalyser, but it is difficult to decipher the traces, as I had major daisy-chaining issues.
Thanks guys!
I've been using the Illumina Truseq small RNA kit, to little success. Starting off with 1ug of total RNA (all with RIN values of 10 I should add!), I took the samples through the whole library prep process, ran them on the polyacrylamide gel (see attached gel pic) and saw faint bands. Unfortunately, when I ran the finished, purified libraries my Bioanalyser results were very substandard (see attached .pdf). Most of them were almost flat lines!! Only one (sample #2) looked vaguely decent (this being the library with the brightest bands on the gel).
I was wondering whether anybody else has had similar problems with this? I have followed the protocol completely, and left the samples to elute for ~18 hours - the protocol says you can leave them in the shaker overnight, so I would expect higher yield if anything!
I also ran the libraries pre-PAGE on the Bioanalyser, but it is difficult to decipher the traces, as I had major daisy-chaining issues.
Thanks guys!
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