TruSeq small RNA

[NicoBxl]

Hi,

Does anybody have already used the TruSeq kit for small-RNA sequencing (miRNA) ?

I heard from people that a lot of reads didn't match with the 3' adapter (36 nt long reads before adapter trimming ).
In summary, they have more than 50% of the reads with no 3' adapter (so RNA longer than 36 nt ).

Does anyone have the same results with this kit ?

Thanks

[cascoamarillo]

Hi Nico,

I can't help you with TruSeq kit. But in my case, my small library (home made 3' and 5' ligation, etc...), after trimming the 3' adapter, shows reads with no adapter sequence (but only 2-3%). But when you look into these reads (in my case 38 nt) you see a bad QC at the 3' end. So maybe that's why the trimming step doesn't detect the adapter.

[epistatic]

I'm about to try the new small RNA kit. Are there any recommendations on how many reads per sample is needed? We have gone to V3 reagents and I am not sure how many I should multiplex in a lane.

[Paul Eijk]

Does someone knows the concentration (units/ul) of the T4 ligase in the miRNA TruSeq kit?
It seems that this item is extact enough for 24 reacties and the only item thats running out as first.

Paul