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Title: Site directed mutagenesis Post by: allan2009 on Oct 29, 2010 Hypothetically i am planning a site-directed mutagenesis experiment and are suppliedwith three sets of primer pairs. The sequences of one primer of each pair are as indicated below:
1. 5'-GAATCTACACACTCGGGGGGGGATCCCCCCCCCCCTAAATCTACC-3' 2. 5'-AACTGGATACACTAGATACATGCATAGAGACATAACTCACTGGGG-3' 3. 5'-ACTAGCCTCAATAGTCAGACTACAACATAGATACAGCATATAATA-3' Which of the three primers would you choose? What reasons for your choice, also why the other two are unsuitable. Title: Re: Site directed mutagenesis Post by: bio_man on Oct 29, 2010 Primer 2 and 3 look very similar, thus you can deduce that they are the unstable ones. I would choose 1 because between guanine and cytosine are bonded together via three bonds whereas thymine and adenine are bonded together by two bonds. Stronger hold equals better results.
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