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Team:EPF-Lausanne/Protocols/Ligation
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==Reaction mix== | ==Reaction mix== | ||
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6:1 molar ratio of insert to vector (~10 ng of vector) | 6:1 molar ratio of insert to vector (~10 ng of vector) | ||
Complete with dH2O up to 10 µl | Complete with dH2O up to 10 µl | ||
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==Molar ratio== | ==Molar ratio== | ||
Revision as of 07:49, 4 September 2009
Contents |
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
