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Wisconsin-Madison/11 June 2009
From 2009.igem.org
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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
