Team:TU Darmstadt/protocols/Dephosphorylation

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<h2><font size="6" color="#F0F8FF" face="Arial regular"> Dephosphorylation </font></h2>
<h2><font size="6" color="#F0F8FF" face="Arial regular"> Dephosphorylation </font></h2>
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Latest revision as of 00:33, 5 October 2013







Lab book | Materials | Protocols

Dephosphorylation

Short explanation
 Antarctic Phosphatase catalyzes the removal of 5' phosphate groups of DNA and RNA and thus prevents religation of cut vectors. It is used before ligation.

Procedure
 The reaction mix contains:
up to 5 µg of digested DNA (no purification needed)
1/10 of reaction volume of 10x Antarctic Phosphatase Reaction Buffer
1 µL of Antarctic Phosphatase

Incubate at 37° C for 30 minutes (3' ends) or 60 minutes (5' ends)
Heat inactivate at 65° C for 15 minutes
Continue with ligation

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