Hello. I need to know the simplest method of doing a point mutation experiment (single-base substitution) on a sample of DNA? I'm very new to this, so the simpler/more well explained, the better.

 

Particularly, I want to perform the mutation to gene trichohyalin-like-1 (TCHHL1). It's on Chromosome I (map position 1q21.3), to change AA to GG (separately) on SNP rs17646946.

 

The TCHHL1 samples I have are 5 shRNA clones of Lentiviral vector type.

 

This is my first experiment with genetic mutation for personal research, so even an outline of a basic method of point mutation in general would be helpful. I'm looking for a more simple tutorial than what I've seen thus far, maybe involving use of loxP? But any method is helpful, really. Thanks in advance.

Edited November 27, 201114 yr by valac

I'd suggest googling some protocol for Site-Directed mutagenesis. (e.g. QuickChange method...)