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Team:HKU HongKong/Data/Protocols.html
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Revision as of 14:25, 23 September 2012
Team:HKU HK
From 2011.igem.org
Molecular Cloning Protocols
Transforming Competent E.coli with the
Plasmid:
Take out the competent E-coli
cells from the -80 freezer. (Keep all tubes on ice).
Add 1uL
of the plasmid DNA in 100uL competent cells (if from Kit). For
transformation of ligated product, add 10uL of the ligated plasmid DNA in
100uL competent
cells.
Incubate on ice for 10 min.
Place
in water bath at 42°C for 90s.
Place
immediately back on ice for at least 2 min.
Add
800uL LB
broth. Incubate for 1hr
at 37°C shaker.
Centrifuge at 130rpm for 5min
to remove supernatant. Re-suspend the pellet in about 10uL of supernatant.
Spread the entire re-suspended
pellet on ampicillin agar dishes.
Incubate for 12-16 hours at 37
°C.
