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	65C for 10 minutes (or 80C)<br>		65C for 10 minutes (or 80C)<br>
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-	<a href = "https://www.neb.com/protocols/1/01/01/dna-ligation-with-t4-dna-ligase-m0202/">DNA ligation with T4 DNA Ligase</a><br>
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Revision as of 03:00, 21 September 2013

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General Ligation Protocol

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)