Team:SCU-China/Linkage

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Latest revision as of 18:55, 17 October 2014

Linkage Protocol

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According to the advice of IGEM Competition, there are two linkage methods that we use in experiment. The first one is “3A” Assembly and the second is “Traditional” Assembly.

For “3A” Assembly, we set up the following system (20 μl):
Linear Backbones from IGEM Competition( with K+, C+, A+, or T+)	About 100 ng
Insert DNA 1	The molar ratio of Insert DNA to Backbone is 3:1 to 5:1
Insert DNA 2	Same to Insert DNA 1
10 x T4 DNA Ligase Buffer	2 μl
T4 DNA Ligase	1 μl
H2O	Up to 20 μl

For “Traditional” Assembly, we set up the following system (20 μl):
Linear Backbones from IGEM Competition( with K+, C+, A+, or T+)	About 100 ng
Insert DNA	The molar ratio of Insert DNA to Backbone is 3:1 to 5:1
10 x T4 DNA Ligase Buffer	2 μl
T4 DNA Ligase	1 μl
H2O	Up to 20 μl

Sichuan university

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