Team:British Columbia/Notebook/Protocols/ligation
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(Created page with "> {{:Team:British_Columbia/Templates/MainHeader}} <html> <div class="container"> <h1>BioBrick Ligation</h1> <br> <h2>Supplies:</h2> <ul> <li>T4 DNA ligase</li> <li>Ligase bu...") |
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<il>*Formula 1: insert mass(ng) = (1/6) x (insert length/vector length) x vector mass (ng)</il> | <il>*Formula 1: insert mass(ng) = (1/6) x (insert length/vector length) x vector mass (ng)</il> | ||
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<il>*Formula 2: volume of insert(µL) = (total plasmid length/ insert length) x (insert mass/plasmid concentration)</il> | <il>*Formula 2: volume of insert(µL) = (total plasmid length/ insert length) x (insert mass/plasmid concentration)</il> |
Revision as of 23:47, 17 October 2014
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BioBrick Ligation
Supplies:
- T4 DNA ligase
- Ligase buffer
- dH2O
- Insert and vector
Steps:
- Get ice.
- Put the reagents on ice.
- Calculate insert vector amount. Add the calculated amounts into eppendorf tube.
- Make a reaction mix with everything but the insert.
- Aliquot the master mix into eppendorf tubes.
- Add insert DNA.
- Let incubate at room temperalerature for at least 2h or overnight.