Wisconsin-Madison/9 July 2009

July 9, 2009
Other:

Advisor Meeting – Brian

Research

ProU, ProV cutting

Ex 20: New Cloning Methods

J. MiniPrep:

1. NudF: Amp

2. Choline: Amp

3. Backbone P1: Amp

K. Digest:

1. NudF – (560+2157=) 2717 bp

2. Choline – (2000+2157=) 4157 bp

3. Backbone P1 – 2157+1069 bp

4. (pVP68K): gsSDMT (Pst1) – (931+5212=) 6143 pb

Cut with (---), Ran on 1% Gel

Left to right:

Uncut: NudF 2B, Choline1, Backbone 1, GSSDMT1, Ladder

Cut: NudF 1A, 1B, 2A, 2B, choline 1,2, Backbone 1, 2, GSSDMT1, 2



Inoculated:

DH1: Blank (no resistance)

Ex 19: Parts from Registry

Transformation plate 1 well O22 into DH10B

Ex 10: Cyanobacteria natural transformation test on 7002 with 1mL culture and 10uL pMMBRraA, allowed to incubate for 24h added 9mL Media A to 7/8 nat trans test cultures and 7.35uL or 10uL of CM, allowed to incubate for 48h made freezer stock of 7002 in 25% glycerol – NP

Primer Design: 1st Antiporter

2nd Antiporter

New ProU

Ex 18: pTaq