different length for primers amplifying - (Mar/14/2008 )
I want to amplify tetR gene from the plasmid, the lengths of forward and reverse primers are different: the forward/45bp reverse/34bp,
Can this pair of primers work?? I do not know, wish to get me answers and explainations. Thanks.
Yes, these can work. Likely the two primers have a 3' end which binds to the template and additional bases (restriction sites etc.) at the 5' end. This 5' end does not play a role during initial binding to the template, but will end up in the final product.
Thanks, I got the target fragments verified by agarose gel electrophoresis, but finally when got the sequencing results, it will be very good.