Team:Newcastle/Ligation

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(New page: {{Team:Newcastle/mainbanner}} For standard ligation 100-200ng of vector DNA is required - can be checked using nanodrop A ligation calcultion can be used however a 1:3 and a 1:5 ratio te...)
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For sticky end ligations incubate on the bench of 3-4 hrs or overnight

Revision as of 13:59, 28 July 2010

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For standard ligation 100-200ng of vector DNA is required - can be checked using nanodrop

A ligation calcultion can be used however a 1:3 and a 1:5 ratio tends to be used instead.

Set up ligation

Reagents 1:3(μl) 1:5(μl) Vector(μl)
V 1 1 1
I 3 5
10*BUFFER 1 1 1
Ligase T4 1 1 1
H2O 4 2 7
Total Volume 10.0 10.0 10.0
For sticky end ligations incubate on the bench of 3-4 hrs or overnight