question related to mutagenesis - (Jan/06/2012 )
i would like to know how many bp we can mute in one mutagenesis? Actually I would like to mute an RNAm in order to make it resistant to siRNA. I've learned that to really disrupt the association RNAm/siRNA, quiet mutations should target the 3' end of the RNAm. I would like to know if several bp can be muted at the same time in one PCR reaction (using one sequence) without disturbing the association of the sequence on the RNAm ( If yes, up to how many? 4 is ok?)
i also would like to know what's the minimum number of muted bp that is sufficient in order to disrupt this assiciation siRNA/RNAm
Thank you for your help.
Yes, you can do more than one base pair at a time, though it gets progressively more difficult as the number gets larger. 4 in a row should be fine.