I have tried to enrich for small RNAs from total RNAs using miRvana kit from Ambion. It definitely got rid of a lot of types of RNAs including abundant rRNAs, but I got enrichment of 90nt size RNAs instead of ~20 to 25nt size small RNAs. Does anyone have any idea of what kinds of RNA this could be? And also can anyone suggest any particular method for enriching small RNAs? I will really appreciate any advice. Thank you very much for all your input in advance. I have attached a summary of what I did and the image of gel after enrichment.
Hi Jeong
Most of the small RNA kits generally give up to around 100 bp size RNA moleculae as you have observed. I have not come across any kit specifically for 20-25 mers...The small RNA preparation can further be resolved on PAGE gel for enrichment...YOu will find protocols in the literture....However, since a lot of other RNA moleculae have already been removed..one can go ahead with Seq and later remove non essential sequences during bioinformatic analysis
bye
If you may try using Norgen Kit. It gives higher RIN value on bioanalyzerfor most of RNA extraction that we tried and successfuly made illumina smallRNA libraries for sequencing. But we gel select small RNA during the protocol and it worked well with good sequencing results..!!
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