primer extension isotopes

Julie Hollien via methods%40net.bio.net (by julie.science from gmail.com)
Tue Sep 25 16:44:01 EST 2007


Dear bionet readers,

I am mapping RNA fragments using primer extension. I am considering using
P-32 end-labeled primers for the primer extension and comparing on the gel
to sequencing reactions using incorporated S-35. Has anyone tried this? Does
anyone foresee a problem with this comparison? Would the products run
differently in a sequencing gel? I am not too concerned about intensity
differences, as my extension products are extremely faint, and I can
sequence off of a cloned full-length template. But I see a potential benefit
to using S-35 as I can store the reactions while trouble-shooting by primer
extension, I've heard that the bands are sharper, and I can minimize my
radiation exposure.

Thanks for any input,
Julie


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