Assembly Related Protocols

From 2008.igem.org

Revision as of 03:04, 2 August 2008 by Sstcheung (Talk | contribs)

Newest EcoRI/BamHI Transfer method (7/31/08)

Digestion:

  • 0.5ul plasmid in entry vector
  • 0.5ul EcoRI
  • 0.5ul BamHI
  • 1ul NEB2 + ATP buffer
  • 7.5ul of water
  • incubate the digestion for 1hr. in the thermocycler, and 20 min for heat kill.

No purification step.

Ligation:

  • 1ul of dilute assembly vector (or 0.5ul if undiluted)
  • 0.5ul ligase

Transform. (same as usual)

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