Wisconsin-Madison/8 July 2009

July 8, 2009
'''Other:

Research Re-Reading Syd’s paper for questions

'''DIFFERENCES IN ISOPENTENOL PRODUCTION

'''Sydnor / IGEM / FIX?

DH1 / DH10B, MG1655, MG1655-DAraBad, DL21-DE3 / Yes

30 C / 37 C (30 C LB) / Yes

5:1 EtAc: Culture / 1:10 EtAc : Culture / Yes

Media see p 6279 / Terrific Broth and LB / ?

Sub Culture / Direct Culture / Yes

MevT (lac) / MevT (ara) / NEED

Carbenicillin / Ampicillin / ?

'''How to put Lac promoter on MevT:

'''Ex 19: Parts from the Registry '''A. MiniPrep:

M12 GFP: amp

gsSDMT: kanamycin

B. Digestion: Ran on 1% Gel

(pSB1A2): M12 GFP (EcoR1) – (876+2079=) 2,952 pb

(pVP68K): gsSDMT (Pst1) – (931+5212=) 6143 pb '''(could not digest, no Pst1)

gssdmt1 gssdmt2, L(1kb), GFP1uncut GFP1cut GFP2 uncut GFP2 cut We took L12 from plate (4713bp) instead of GFP in M12 Need to plate 1.M12 and digest gsSDMT

'''Ex 20: New Cloning Methods

H. Growth – Plates

1. NudF: (2white, 0red)

2. Choline: (1white, 12red)

3. Backbone P1: (1red, 1white)

I. Inoculate (10ml LB, 10uL Amp)

1. NudF: Amp

2. Choline: Amp

3. Backbone P1: Amp

'''Ex 10: Cyanobacteria natural transformation test on 7002 with 1mL culture and 1uL pMMBRraA, allowed to incubate 24h inoculated new 7942 and 7002 liquid cultures in appropriate media, aerated