"
User:University of Washington/5 September 2008
From 2008.igem.org
(Difference between revisions)
(→LuxR from AraC and TetR(Faifan)) |
|||
| (2 intermediate revisions not shown) | |||
| Line 18: | Line 18: | ||
!0%, 0 ul!!0.05%, 1ul!!0.1%, 2ul!!0.50%, 10ul!!1.00%, 20ul | !0%, 0 ul!!0.05%, 1ul!!0.1%, 2ul!!0.50%, 10ul!!1.00%, 20ul | ||
|- | |- | ||
| - | !rowspan=5 valign=center|aTc(ng/ml), 10ug/ml-aTc added(ul)!!0, 0ul|||||||||| | + | !rowspan=5 valign=center|aTc(ng/ml), 10ug/ml-aTc added(ul)!!0, 0ul||A1||A2||A3||A4||A5 |
|- | |- | ||
| - | !50, 1ul|||||||||| | + | !50, 1ul||B1||B2||B3||B4||B5 |
|- | |- | ||
| - | !100, 2ul|||||||||| | + | !100, 2ul||C1||C2||C3||C4||C5 |
|- | |- | ||
| - | !500, 10ul|||||||||| | + | !500, 10ul||D1||D2||D3||D4||D5 |
|- | |- | ||
| - | !1000, 20 ul|||||||||| | + | !1000, 20 ul||E1||E2||E3||E4||E5 |
|} | |} | ||
*added dH2O to final the volume to 200 ul | *added dH2O to final the volume to 200 ul | ||
| Line 32: | Line 32: | ||
*set the machine to run 24 hrs, read every 10 mins | *set the machine to run 24 hrs, read every 10 mins | ||
| + | == Conjugation (Scott)== | ||
| + | [[Team:University_of_Washington/Protocols#Method_.232| Conjugation method #2]]<br> | ||
| + | ·pAC99 + pUB307 | ||
| + | [[UW Glycerol Stocks| Glycerol Stock]]<br> | ||
| + | ·pAC99<br> | ||
| + | ·pAC99 + pUB307 | ||
---- | ---- | ||
Back to [[Team:University_of_Washington/Notebook#Notebook]] | Back to [[Team:University_of_Washington/Notebook#Notebook]] | ||
Latest revision as of 19:36, 10 September 2008
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
Back to Team:University_of_Washington/Notebook#Notebook
