User:University of Washington/15 July 2008

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RP4 Conjugation

Mini prep
·pAC88

Bac-Bac conjugation protocol #1
·DH5alpha + RP4
·DH5alpha + pCS26-pac

LuxR from AraC and TetR

- Miniprepped 12 overnight culture of mutated AraC into 30 ul EB

- Ordered sequences of all 12: 8 ul (1 umol/mL) forward primer + 4 ul DNA

- Grew 4 overnight culture of D29 from Elowitz's lab

LuxR from pLac

- Parts R0010, E0420, and I0462 were digested, ran on gel, then extracted for assembly of two new parts: R0010 in front of E0420, and R0010 in front of I0462. Biocircuits lab protocol used.

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