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Team:Washington/GENERAL LIGATION PROTOCOL
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Revision as of 06:26, 7 September 2013
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Reaction Mixture:
7uL insert (1:3 to 1:7 ratio of vector:insert) & (2:6) also works well
1uL vector
1uL T4 ligase buffer
1uL T4 ligase
10ul total
Incubate at 16C overnight, or room temperature for 30 minutes to 1 hour.
Heat inactivation after ligation, before transformation can increase transformation efficiency.
65C for 10 minutes (or 80C)
DNA ligation with T4 DNA Ligase
