Mutagenesis - To amplify fragments for mutagenesis (Nov/21/2008 )
I have one question regarding to mutagenesis. If the size of fragment 1 is very short about 45 bp, how do we can make mutagenesis, seems it's not possible to obtain the small size, unless if we got another way such as replacement of primer to pET Near up to obtain about 100bp (45bp + about 55 bp of pET Near Up) I guess not really sure... :huh
For fragment 2, it's ok about 500 bp..
I need answer ASAP
Get 2 primers that would essentially cover the whole length of the fragment between them but that only overlap in the middle ie the 3' end of your left primer overlaps the 3' end of your right primer, then just PCR them together and you will get a fragment of DNA 45bp long, exactly as you like it.