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	=General Ligation Protocol=		=General Ligation Protocol=

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Revision as of 20:47, 13 June 2012

General Ligation Protocol

1. Ligate the Upstream and Downstream Parts into the digested Destination Plasmid.

   Upstream Part digestion:	   2 µl

   Downstream Part digestion:	   2 µl

   Destination Plasmid digestion: 2 µl

   10X T4 DNA Ligase Buffer:	   2 µl

   T4 DNA Ligase:	           1 µl

   H2O:	                  11 µl

1. Incubate at room temperature for 10 minutes and then heat inactivate at 80°C for 20 minutes.

1. Transform 2 μl of the ligation product into 50 μl of competent E. coli cells (or other suitable host strain). Select using the antibiotic corresponding to the Destination Plasmid.