Team:EPF-Lausanne/Protocols/Ligation

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Revision as of 07:49, 4 September 2009

Contents

Ligation Protocol





Reaction mix

T4 ligase buffer 1 µl
T4 ligase 0.5 µl


6:1 molar ratio of insert to vector (~10 ng of vector) Complete with dH2O up to 10 µl

Molar ratio

insert mass ng= 6 ∙ insert lengthvector length ∙vector mass [ng]


Notes

In general, we can take 1 µl of vector for 3 µl of insert.

To control ligation specificity, you need to have a control with the vector only.


Incubation

30 min at RT