Team:Brown/Protocols/Ligation

From 2010.igem.org

Ligation

  • 7.5 µl 2X DNA ligase buffer (rapid)
  • 1 µl T4 DNA ligase
  • 0.5 µl vector
  • 1.5 µl insert
  • 4.5 µl dH2O

15 µl total volume

24-36 hours at 4°C. Alternatively 1 to 2 hours at room temperature may also work, although we found a generally lower ligation efficiency using this method.

Note: Generally aim for a 1:3 vector to insert ratio.

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