Dear all,
I have generated Stable Cell lines via G418 selection.
I have used pCMV-miR vector.Except Neo-GFP region, my chance is 2/5 to integrate genome without damaging CMVpromotor-insert-PolyA tail region.
So my question is that;
Can I try to detect PolyA tailed pri-miRNA forms by oligodT cDNA synthesis. Due to overexpression, as far as I am concerned I should.
and the second question is that, if my PolyA signal is destructed during integration, ( I can check via genomic pcr CMVpromotor to insert) can this destruction effect miRNA expression? Normally miRNAs do not require polyA signal for biogenesis but they require correct transcriptional termination.
Hey,
They are Stem-loop primer cDNA synthesis based. I am looking for pri-miRNA forms which are poly adenilated.
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