Team:Macquarie Australia/Protocols/ligations

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<p>The mixtures were incubated for 10 minutes and then heat inactivated at 80&deg;C for 20 minutes.</p>
<p>The mixtures were incubated for 10 minutes and then heat inactivated at 80&deg;C for 20 minutes.</p>
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<p>2&micro;L  of the ligation product was transformed into 50&micro;L  of competent <i>E. coli</i>, the transformants were then incubated overnight.</p>
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<p>2 &micro;L  of the ligation product was transformed into 50 &micro;L  of competent <i>E. coli</i>, the transformants were then incubated overnight.</p>
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Revision as of 07:17, 23 September 2012



Ligation Procedure

A reaction mixture was prepared containing,

Element Volume
Upstream Digestion part 2 µL
Downstream Digestion part 2 µL
Destination Plasmid 2 µL
10X T4 DNA ligase buffer 2 µL
T4 DNA ligase 1 µL
H2O 11 µL

The mixtures were incubated for 10 minutes and then heat inactivated at 80°C for 20 minutes.

2 µL of the ligation product was transformed into 50 µL of competent E. coli, the transformants were then incubated overnight.