Originally posted by linda
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I've seen protocols that "force clone" the 3' ends of cDNAs away from the sequence priming site. If you like oligo dT methods, I would recommend one of those. (In brief, the one I've seen uses only T4 polymerase to blunt after nebulization--no T4PNK. If you add only PA adaptor to the ligation it forces ligation to the broken end because the cDNA adaptor end is not phosphorylated. The PB adaptor is added subsequently added by "step out" PCR, using PA and PB+SMART-CAP primers.)
Originally posted by linda
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Phillip
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