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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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N-terminal deletion primers
Posted by: jbwaj (IP Hidden, New member, 3)
Date: December 7, 2005 02:50PM
Hi all
I need to construct a N-terminal deletion of a protein through PCR. I was told that the best way to do it is to amplify the C-terminal region with a Nde1 and BamH1 recognition site and insert it into a suitable vector. I have a couple of questions regarding the primer design: I assume that there has to be an ATG codon just before the C-terminus so that the translation starts there.Am I right?? Also, the BamH1 recognition site is GGATCC....will this additonal insertion in the primer affect its ability to anneal to the parent template? Thanks!! P.S: I am not a molecular biologist and hence not too familiar with PCR methods.
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