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Team:Washington/Gram Positive/Build
From 2010.igem.org
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=Mutate DNA= | =Mutate DNA= | ||
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OUTLINE | OUTLINE | ||
Revision as of 23:06, 7 September 2010
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
Transform E. coli with the mutant plasmid
Grow and OD until desired OD is reached
Induce bacteria to produce our protein
Grow for 24 hours
Harvest Protein
Spin down cells
Lyse cells
Purify cells
concentrate as needed
