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How to do a Point Mutation?


valac

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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 by valac
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