pcr cloning blues

Rory O'Brien robrien at ollamh.ucd.ie
Thu May 18 14:32:22 EST 1995


If any can shed light on this problem I would be pleased. The problem is that while attempting to PCR amplify certain gene sequences from a cDNA library I have succeeded in amplifying fragments that correspond sizewise with the desired product. These fragments however, are seen to be different entities when sequenced e.g,cloning vector sequences. This has been observed over multifarious 'optimized'reaction conditions. Why is this?, I ask myself. I also have problems nesting certain southern identified products where fragments much smaller appear, also under a range of conditions. I have tried every polymerase under the sun! Suggestiions PLEASE!




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