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miRNA seq info

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#1 leo77



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Posted 09 March 2011 - 02:41 PM

Dear all,
I'm dealing with this problem and I'd really like to have input from you. I'm working on a miR and found out that using a commercial probe I can detect a band by northen blot much higher than the one at 22 expected. The band I saw is higher than 100bp and based on my experience is not the pre-miR. My aim now is to find out what that is, and I'm working on a protocol to sequence it. My idea was to hybridize in vitro the total RNA with a biotynilated probe and then use a streptavidine-magnetic beads in order to purify the complex. At this point I thought I can generate cDNA and then seq it designing a complementary primer on the miR sequence, but not sure if that could be the best idea. Any suggestion from you guys will be very important to me.

Thanks to all


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