Léa
Glycerols and minipreps
- wRBS/LacZα
- sRBS/LacZα
- LuxI
- pLux
- attC/t/attC/mRFP
- Pm/mRBS/LuxR
- Pm/wRBS/LuxR
- wRBS/TetA(C)/LuxI
- mRBS/TetA(C)/LuxI
- sRBS/TetA(C)/LuxI
- Pm/wRBS/LuxR/attC/t/attC/mRFP
- pLux/LuxI (started with 3 colonies)
Nanodrop of the minipreps
Sample name |
260/280 |
230/280 |
ng/µl
|
|
|
|
|
|
|
|
|
wRBS/LacZα |
1.92 |
2.34 |
128.7
|
sRBS/LacZα |
1.91 |
1.70 |
175.1
|
pLux |
1.90 |
2.07 |
222.9
|
attC/t/attC/mRFP |
1.88 |
1.91 |
219.0
|
Pm/mRBS/LuxR |
1.93 |
2.35 |
686.8
|
Pm/wRBS/LuxR |
1.87 |
2.15 |
437.8
|
wRBS/TetA(C)/LuxI |
1.96 |
2.27 |
235.4
|
mRBS/TetA(C)/LuxI |
1.95 |
2.19 |
282.6
|
sRBS/TetA(C)/LuxI |
1.91 |
2.26 |
340.2
|
Léa
Restriction digest with SpeI and PstI : 20µl miniprep + 2x1µl enzyme + 2.5µl B2 + 0.25µl BSA
- wRBS/LacZα
- sRBS/LacZα
- pLux
- Pm/mRBS/LuxR
- Pm/wRBS/LuxR
- Pm/wRBS/LuxR/attC/t/attC/mRFP (5µl miniprep + 2x1µl enzyme + 2.5µl B2 + 0.25µl BSA + 15µl water)
Restriction digest with XbaI and PstI :
- LuxI (20µl miniprep + 2x1µl enzyme + 2.5µl B3 + 0.25µl BSA)
- attC/t/attC/mRFP (20µl miniprep + 2x1µl enzyme + 2.5µl B3 + 0.25µl BSA)
- pLux/LuxI, 3 different minipreps (5µl miniprep + 2x1µl enzyme + 2.5µl B3 + 0.25µl BSA + 15µl water)
Aleksandra
Gel electrophoresis
- wRBS/TetA(C)/LuxI (PCR of the uncut plasmid)
- mRBS/TetA(C)/LuxI (PCR of the uncut plasmid)
- sRBS/TetA(C)/LuxI (PCR of the uncut plasmid)
- Ladder 1kb
- wRBS/TetA(C)/LuxI (PCR of the plasmid cut by AlWNI), 2 wells
- -
- mRBS/TetA(C)/LuxI (PCR of the plasmid cut by AlWNI), 2 wells
- -
- sRBS/TetA(C)/LuxI (PCR of the plasmid cut by AlWNI), 2 wells
File:Gel1608 1.jpg
File:Gel1608 1cut.jpg
Gel extraction
- wRBS/TetA(C)/LuxI (PCR of the plasmid cut by AlWNI)
- mRBS/TetA(C)/LuxI (PCR of the plasmid cut by AlWNI)
- sRBS/TetA(C)/LuxI (PCR of the plasmid cut by AlWNI)
Nanodrop of the gel extractions
Sample name |
260/280 |
230/280 |
ng/µl
|
|
|
|
|
|
|
|
|
wRBS/TetA(C)/LuxI |
1.91 |
0.32 |
47.9
|
mRBS/TetA(C)/LuxI |
1.87 |
0.25 |
40.7
|
sRBS/TetA(C)/LuxI |
1.87 |
0.35 |
63.0
|
Restriction digest of the with XbaI and PstI : 20µl miniprep + 2x1µl enzyme + 2.5µl B3 + 0.25µl BSA
- wRBS/TetA(C)/LuxI
- mRBS/TetA(C)/LuxI
- sRBS/TetA(C)/LuxI
PCR kit purification, elution in 35µl warm water
Sample name |
260/280 |
230/280 |
ng/µl
|
|
|
|
|
|
|
|
|
wRBS/TetA(C)/LuxI |
1.68 |
1.40 |
19.8
|
mRBS/TetA(C)/LuxI |
1.59 |
0.96 |
21.6
|
sRBS/TetA(C)/LuxI |
1.73 |
1.54 |
28.2
|
pLux |
1.75 |
1.37 |
62.7
|
Ligations
- 1)Pm/sRBS/LuxR + wRBS/TetA(C)/LuxI (0.5µl + 4.5µl + 0.5µl ligase + 1µl buffer + 3.5µl water)
- 2)Ps/wRBS/LuxR + wRBS/TetA(C)/LuxI (1µl + 4.5µl + 0.5µl ligase + 1µl buffer + 3µl water)
- 3)Ps/mRBS/LuxR + wRBS/TetA(C)/LuxI (1µl + 4.5µl + 0.5µl ligase + 1µl buffer + 3µl water)
- 4)Ps/sRBS/LuxR + wRBS/TetA(C)/LuxI (0.5µl + 4.5µl + 0.5µl ligase + 1µl buffer + 3.5µl water)
- 5)Pm/sRBS/LuxR + mRBS/TetA(C)/LuxI (0.5µl + 4.5µl + 0.5µl ligase + 1µl buffer + 3.5µl water)
- 6)Ps/wRBS/LuxR + mRBS/TetA(C)/LuxI (1µl + 4.5µl + 0.5µl ligase + 1µl buffer + 3µl water)
- 7)Ps/mRBS/LuxR + mRBS/TetA(C)/LuxI (1µl + 4.5µl + 0.5µl ligase + 1µl buffer + 3µl water)
- 8)Ps/sRBS/LuxR + mRBS/TetA(C)/LuxI (0.5µl + 4.5µl + 0.5µl ligase + 1µl buffer + 3.5µl water)
- 9)Pm/sRBS/LuxR + sRBS/TetA(C)/LuxI (0.5µl + 4.5µl + 0.5µl ligase + 1µl buffer + 5µl water)
- 10)Ps/wRBS/LuxR + sRBS/TetA(C)/LuxI (1µl + 4.5µl + 0.5µl ligase + 1µl buffer + 4.5µl water)
- 11)Ps/mRBS/LuxR + sRBS/TetA(C)/LuxI (1µl + 4.5µl + 0.5µl ligase + 1µl buffer + 4.5µl water)
- 12)Ps/sRBS/LuxR + sRBS/TetA(C)/LuxI (0.5µl + 4.5µl + 0.5µl ligase + 1µl buffer + 5µl water)
Controls:
- 13)Pm/sRBS/LuxR (0.5µl + 0.5µl ligase + 1µl buffer + 8µl water)
- 14)Ps/wRBS/LuxR (1µl + 0.5µl ligase + 1µl buffer + 7.5µl water)
- 15)Ps/mRBS/LuxR (1µl + 0.5µl ligase + 1µl buffer + 7.5µl water)
- 16)Ps/sRBS/LuxR (0.5µl + 0.5µl ligase + 1µl buffer + 8µl water)
Raphaël
Glycerols
- Ps/sRBS/LuxR/attC/t/attC/mRFP and pBAD/IntI, 4 cultures
- Ps/wRBS/LuxR/attC/t/attC/mRFP and pBAD/IntI, 4 cultures
- Ps/mRBS/LuxR/attC/t/attC/mRFP and pBAD/IntI, 4 cultures
- Pm/sRBS/LuxR/attC/t/attC/mRFP and pBAD/IntI, 4 cultures
- pLux/RBS/LuxI, 3 cultures
- Pm/wRBS/attC/t/attC/mRFP, 1 cultures
Minipreps
- LuxI
- Pm/wRBS/LuxR/attC/t/attC/mRFP
- pLux/LuxI, 3 cultures
Nanodrop of the minipreps
Sample name |
260/280 |
230/280 |
ng/µl
|
|
|
|
|
|
|
|
|
LuxI |
1.97 |
2.25 |
244.2
|
Pm/wRBS/LuxR/attC/t/attC/mRFP |
1.96 |
2.27 |
256.8
|
pLux/LuxI cult1 |
1.96 |
2.12 |
195.1
|
pLux/LuxI cult2 |
2.60 |
2.27 |
186.2
|
pLux/LuxI cult3 |
2.00 |
2.16 |
174.0
|
Overnight cultures
- wRBS/LacZα/LuxI, 4 colonies
- sRBS/LacZα/LuxI, 4 colonies
- LuxI
|