I plan to cut out a fragment from a plasmid and then amplify this fragment with PCR.
In addition to amplify the fragment, i also want to add something behind this fragment to modify it. and then ligate it to another plasmid.
I read a paper that used a 42bp forward primer and a 106bp reverse primer (BMC Biology 2008, 6:40).
My fragment is 2kb long and the fragment that i want to add behind it is 64bp long. So i designed a 47bp forward primer and a 96bp reverse primer based on this paper.
I am new in the field, but i always heard people say that ~20 to 25bp primers are optimal. SO, i was wondering is it possible to use such long primers to PCR my fragment?? How will it work?? is there any specific point that i have to pay attention while doing long primers PCR???
thank you in advance for your suggestion.
Edited by Ah-Do, 17 March 2009 - 05:25 AM.