bchs1b@jetson.uh.edu (09/24/90)
I need to make some site directed insertions of about 8-10 bases. I have normally been doing sited directed mutagenesis using Kunkel method (dut ung M13 template) with good success, but have never tried insertions. How many bases on either side of the insertion do I need to make it work well, assuming reasonable GC distribution. Any problems to watch out for? Second related question, are there any procedures for doing site directed mutagenesis on double stranded plasmids which are as easy and efficient as the Kunkel method? Or is it worthwhile cloning things into M13 everytime (I have reasons not to use filamid vectors like pEMBL and pUC118). thanks, mike benedik biochemical and biophysical sciences university of houston benedik@uh.edu or benedik@uhou