IGEM:Harvard/2010/General Protocols/BioBricks

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A Biobrick is a piece of DNA flanked by a STANDARD set of restriction sites: EcoRI (E), NotI (N), XbaI (X) upstream to the left; SpeI (S), NotI (N), PstI (P) downstream to the right.

  • The sequence of the BioBrick cloning site is: gaattc(E) gcggccgc(N) tctag(X) [BioBrick Part] actagt(S) gcggccg(N) ctgcag(P)
  • E-N-X are referred to as the "BioBrick Prefix" and S-N-P are referred to as the "Biobrick Suffix"
  • The BioBrick part itself must NOT contain any of the standard restriction sites, or else it will be chopped up into pieces during assembly
  • The Biobrick vector we will be using is called V0120, a high-copy plasmid (~300 ng/uL from a mini-prep of a 5 mL culture)