Is there any easier way to make deletion and site-mutation at same time? - (Sep/13/2006 )
I have a luc reporter construct containing 300bp promoter, and I made one site and double sites mutation using QuikChange multi site-directed mutagenesis kit. It is very perfect. However, now
I would like to make the same constuct only containing 250bp promoter, namely, delete 50bp, at same time make single site mutation. Could I do the deletion and mutation in one step? the mutation site is far away from the deletion squence. What should I do? Is there anybody know answers? Thanks
not too sure about really understand where you want the "second class" mutations to be, but is it possible to simply PCR the promoter with the first round of mutations and clone it?
If not, please explain little more your exp.
may you tell me where are the mutations in the first assay ? and relevant restriction sites in the area of the 5' and 3' of your promoter...
Thank yu Fred 33, I have know how to do it. Thank you so much
well, it's really amazing to have helped without knowing the whole stuff...