Team:Newcastle/Ligation

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(Set up ligation)
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=Ligation=
For standard ligation 100-200ng of vector DNA is required - can be checked using nanodrop
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.  
A ligation calcultion can be used however a 1:3 and a 1:5 ratio tends to be used instead.  
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===Set up ligation===
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=Procedures=
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Set up the ligation mix according:
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For sticky end ligations incubate on the bench of 3-4 hrs or overnight
For sticky end ligations incubate on the bench of 3-4 hrs or overnight
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Revision as of 15:08, 28 July 2010

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Ligation

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.

Procedures

Set up the ligation mix according:

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

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