Wisconsin-Madison/11 June 2009

June 11, 2009
Other: Meeting with Brian (Notes – iGEM: 0 Summer Meeting)

Ex 10:	Made Terrific Broth for triple transformed cell medium

Inoculation of triple transformation

Induction: 	1. Irrabinose -  0.8 – 1% g/100mL

2. Wait 30 minutes

3. 1mM conc. IPTG (100uL/100mL)

4. Layer of Dodecane to trap isopentenol

Ex 11: Salt tolerance of E.coli DH10B:

Use plate reading machine to chart OD over time

ExA: NaCl 0-1 mol in one tenth intervals (NaCl - Mm 58.44) in Liquid salt-free LB 30uL of DH10B ecoli, 200uL of LB OD taken every 30 min for 48 hours

Conc/NaCl (g)/Actual (g)

0.0M/0.0/0.0

0.1M/ 0.116886/ 	0.1157

0.2M/ 	0.233772/ 	0.2355

0.3M/ 	0.350658/ 	0.3560

0.4M/ 	0.467544/ 	0.4633

0.5M/ 	0.584430/ 	0.5859

0.6M/ 	0.701316/ 	0.7034

0.7M/ 	0.818202/ 	0.8156

0.8M/ 	0.935088/ 	0.9348

0.9M/ 	1.051974/ 	1.0577

1.0M/ 	1.16886/ 	1.1692

Inoculation of DH10B to run experiment tomorrow

Ex 9: Transformation of spinach cDNA into E.Coli DH10B

Add 50uL of to ~100ug precipitated DNA from Mike Ziemak

(pREP3-A/spinach PEAMT DNA)

Ex 13 Design: Thermocycles for Choline, Pro U, YhfR, NudF

- inoculated cultures of pSB1A3, pRL1383A, and pMMBRraA in LB media with appropriate antibiotics to remake freezer stocks - NP - inoculated liquid culture of pNudF, pMBI, pMevT transformed e coli in LB with appropriate antibiotics to test isopentenol production by induction tomorrow - NP