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Old 11-09-2013, 05:19 AM   #5
Location: Madrid

Join Date: Dec 2011
Posts: 13

Originally Posted by frang11 View Post
Thanks a lot Kcchan. The gel purification was our idea at the beginning but at the senquencing service of the University of Laval, they told us was unnecessary. I will reset my protocol as soon as possible to have better result. Do you have any idea why in our samples we have a shift in the main band? why the main peak is at 153? Could it be an effect of the low quality samples?
Sorry for asking a lot of question but we would really like figure out every details
I agree with Mcchan in that the best method for size selection is the gel cut. We have obtained very good results both with Truseq and NEBnext. However the size that you can be obtained depend of nature of sample (about 145 pb miRNAs and 155 pb piwi RNAs). I suggest you see the TruSeqSmallRNA Sample PreparationGuide pages 27-28. In attachment you can see our last library prepared with NEBnext.
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