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miRNA seq info


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#1 leo77

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

Leo




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