Team:ESBS-Strasbourg/Ligation

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Revision as of 07:59, 5 August 2008 by KatjaK (Talk | contribs)

1)Remarks:



2)Protocol:

  • For insert/vector ligation:

[http://openwetware.org/wiki/DNA_Ligation see iGEM protocol]

  • For intra vector ligation:

5 T4 DNA ligase unit, for 25-50ng of DNA

  • 2c)
    • 50-400 ng vector
    • 1 to 3 times the molar amount of the insert
    • 2 µl ligation buffer
    • 1 µl ligase
    • to 20 µl water

-> vortex and spin down
-> 1h at 22°C water bath (in 4°C room)
-> enzyme inactivation at 65°C for 10 min

Protocols