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User:University of Washington/5 September 2008
From 2008.igem.org
LuxR from AraC and TetR(Faifan)
- measured Absorbance of the overnight culture at 660 nm = 1.5
- diluted 60 ul culture into 5.40 ml Tsy+KAN. Absorbance at 660 nm = 0.035
- incubated 37 degree Celsius for 2.5 hrs, Absorbance at 600 nm = 0.7
- diluted 1 ml culture into 5 ml Tsy+Kan, OD@600nm = 0.165
- set up 96 well plate reader experiment:
- 5x5, 160 ul culture
| Arabinose(%), 10%Arabinose added (ul) | ||||||
|---|---|---|---|---|---|---|
| 0%, 0 ul | 0.05%, 1ul | 0.1%, 2ul | 0.50%, 10ul | 1.00%, 20ul | ||
| aTc(ng/ml), 10ug/ml-aTc added(ul) | 0, 0ul | A1 | A2 | A3 | A4 | A5 |
| 50, 1ul | B1 | B2 | B3 | B4 | B5 | |
| 100, 2ul | C1 | C2 | C3 | C4 | C5 | |
| 500, 10ul | D1 | D2 | D3 | D4 | D5 | |
| 1000, 20 ul | E1 | E2 | E3 | E4 | E5 | |
- added dH2O to final the volume to 200 ul
- added 40 ul of mineral oil to each well to protect evaporation
- set the machine to run 24 hrs, read every 10 mins
Conjugation (Scott)
Conjugation method #2
·pAC99 + pUB307
Glycerol Stock
·pAC99
·pAC99 + pUB307
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