User:University of Washington/10 July 2008

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Contents

RP4 Conjugation

Conjugation protocol #1
·DH5α
·DH5α + S17-1 w/ RP4
·DH5α + naked RP4 DNA
·DH5α + amp resistant RFP plasmid

Lambda Red

·Synthesis tetR insert for KorA and trbA replacement

non-RP4 Conjugation

Mini Prep & Glycerol Stock
·CV13(Yep13)
·pDPT51

LuxR from AraC and TetR

- GFP E0240 grew, got some single colonies, stored in a fridge.

- Finished up QuikChange Mutagenesis: Dpn1 digestion.

- Did PCR purification with 30 ul EB Buffer.

- Transformed mutated AraC plasmids into XL1 Blue cells.

//Note: Electroporator 200 Ohms, 1.5 kV, with Tsy Media
//Note: Failed to put cells on ice while bringing them over to do the electroporation. Hopefully, they will still grow.

    • Tube 1: 45 ul XL1 Blue + 15 ul DNA#1
    • Tube 2: 45 ul XL1 Blue + 2 ul DNA#1
    • Tube 3: 45 ul XL1 Blue + 15 ul DNA#2
    • Tube 4: 45 ul XL1 Blue + 2 ul DNA#2
    • Tube 5: 45 ul XL1 Blue

- Plated 200 ul each on LB agar + Amp


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