Wisconsin-Madison/29 June 2009

June 29, 2009
'''Ex 16:

MiniPrep: of P2 colonies A-C, ProU colonies A-3, NudF colonies A-3, GFP+ProU

'''Inoculate Cultures: YhfR, BB (P1)

U   U   U    F   F   F   B*  B*  B*UG  L   U   F    B* UG	Left of ladder-cut, right of ladder-uncut Expected sizes: F:2700bp, U:2350bp, B*:2150bp, UG:2650bp

'''Ex 17: GFP Regulation by ProU Promoter

'''Experimental Design:

0.0-1.0M in increments of 0.2 concentrations of (4 wells of each concentration) 1. Wild Type E.Coli 2. GFP only 3. ProU + GFP

''(Plan to include the SAM synthase (METK) to boost gsSDMT function, and eventually getting rid of the GFP and evaluating solely on growth)

''4. ProU + GFP + gsSDMT ''5. ProU + GFP + gsSDMT + MetK

6. Salt Free LB – start (2 wells of each concentration) 7. Water (2 wells of each concentration)

1. Grow four different cultures till OD600=0.4 to 0.6 2. Induce cultures with various concentrations of salt in the plate reader 3. Let the plate reader shake and incubate overnight 4. Take OD and excitation/emission value at 501/511nm during the entire time course

''Test 4:

'''Conc of salt (M): 0.0 / 0.2 / 0.4 / 0.6 / 0.8 / 1.0

DH10B

GFP only ProU+GFP

Salt Free LB

Water

''0D 600

.401	GFP+ProU

.425	GFP

.415	DH10 B

- Incubator overnight (37 C)

'''Ex 10: Cyanobacteria Cyano antibiotic test Made 10:1 dilutions of 6803 to achieve 110 mL cultures, doubled the volume of transformed cells, made freezer stocks of extremely dark green 7942, tested antibiotics and other things with 7942 - NP