Mutate DNA

Image of kunkels overview, all steps in image format

OUTLINE

Order Olglionucleotides with the desired mutation

Create ssDNA of wild type gene

Allow Olglios to annele to the ssDNA

Use polymerase and DNTPs to synthesize the rest of the gene

Grow Protein

Overview

Transform E. coli with the mutant plasmid

Grow and OD until desired OD is reached

Induce bacteria to produce our protein

Image of lac operon

Grow for 24 hours

Harvest Protein

Overview

Spin down cells

Lyse cells

Purify cells

Image of how it bonds to the beads

concentrate as needed