Cloning IPCR Products

Stephen Dibley stephendibley at ozemail.com.au
Thu Aug 30 21:54:01 EST 2001


Does anyone have any experience with cloning inverse PCR products? I
generated two fragments via inverse PCR and cloned them into the cloning
vector pDrive (Qiagen). Digestion analysis of these plasmids shows that
they contain the correct size insert, but PCR analysis on these plasmids
always produces multiple bands. This occurs with both sequencing primers
(SP6 and T7) and insert specific primers. I have increased the annealing
temperature as still recovered two bands. This problem is also stopping
my from sequencing the insert. Any suggestions on what to do would be
appreciated.

Thanks in advance,

Stephen




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