Wisconsin-Madison/9 July 2009

From 2009.igem.org

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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

0706 gel.jpg 0706 ladder.jpg


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