Oligo directed Insertions???

[bchs1b at jetson.uh.edu]

	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 at uh.edu or benedik at uhou