IGEM:Harvard/2010/General Protocols/BioBricks

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.

• Standard Assembly of BioBricks is explained on the Parts Registry Website

• 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)