Team:Macquarie Australia/Protocols/Ligation

From 2013.igem.org

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<h1>Ligation Procedure</h1>
<h1>Ligation Procedure</h1>
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A reaction mixture was prepared containing,
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<b>Prepare</b> the following reaction mixture
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<thead><tr><th>Element </th><th>Volume</th></tr></thead>
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<tbody><tr><td>Upstream Digestion part </td><td>2 µL</td></tr>
<tbody><tr><td>Upstream Digestion part </td><td>2 µL</td></tr>
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Incubate each ligation mix at 30°C for 30 minutes, followed by heat inactivation at 80°C for 20 minutes.
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We incubated each ligation mix at 30°C for 30 minutes, followed by heat inactivation at 80°C for 20 minutes.
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transform 4 µL of the ligation product into 100 µL of competent E. coli, then incubate the transformants for one hour.
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4 µL of the ligation product was transformed into 100 µL of competent E. coli, the transformants were then incubated for one hour.
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[[File:LigationQLD.jpg|500px|thumb|center|Ligation]]

Latest revision as of 07:18, 13 September 2013


Ligation Procedure


Prepare the following reaction mixture

Element Volume
Upstream Digestion part 2 µL
Downstream Digestion part2 µL
Destination Plasmid1 µL
10X T4 DNA ligase buffer2 µL
T4 DNA ligase1 µL
H2O12 µL

Incubate each ligation mix at 30°C for 30 minutes, followed by heat inactivation at 80°C for 20 minutes.

transform 4 µL of the ligation product into 100 µL of competent E. coli, then incubate the transformants for one hour.


Ligation
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