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Scripps Laboratories for tRNA Synthetase Research 

Protocols

Competent Cell Transformation

Protocol for ampicillin transformation:

Pre-warm an ampicillin plate at 37 degrees

Thaw an aliquot of XL-10 competent cells

Add plasmid and gently mix (keep volume of added plasmid <5% of total reaction volume)

Incubate on ice for 5-10 minutes

Spread this reaction on your warm ampicillin plate

All finished!

 

Protocol for kanamycin & other antibiotic selection plates:

 Pre-warm your plate at 37 degrees

Thaw an aliquot of XL-10 competent cells

Add plasmid and gently mix (keep volume of added plasmid <5% of total reaction volume)

Incubate on ice for 5-10 minutes

Add 4x volumes of LB/SOC media to your reaction mixture

Transfer this mixture to a culture tube and rotate at 37 degrees for 1 hour

Spread this reaction on your warm antibiotic plate

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