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Rapid DNA Ligation

Copyright 2012 Molecular Station

If you want quick, fast or rapid methods of DNA ligation for your cloning then you can use the:

Quick DNA Ligation Protocol

  • Use 8 ul of insert and vector (less than 200 ng)
  • It is suggested by BMC to use 3:1 vector to insert ratio if vector is de-phosphorylated
  • Use 2 ul of 5x DNA dilution buffer (Tube #1)
  • Add 10 ul of 2X ligation buffer (Tube #2)
  • Add 1 ul of ligase (assume you have to use "special ligase" Tube #3)
  • 5 min at room temp

Special Rapid DNA Ligases Include:

NEB's Quick Ligation Kit (5 Minute ligation cloning)

 

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