August/9 October 2009
From 2009.igem.org
We collected sequence data from the automated sequencer and used sequence alignment software to check the sequences against those obtained from the Parts Registry. The results were as follows:
Senders:
LuxI - mismatched
LasI - OK
RhlI - OK
CinI - unconfirmed (DNA sample concentration too low?)
Receivers:
LuxR - OK
LasR - mismatched
RhlR - OK
CinR - unconfirmed (DNA sample concentration too low?)
The DNA for Cin signaling system will be transformed, amplified and purified to obtain at higher concentration and PCR sequencing repeated on them. As time is running out, for the final circuit we shall use parts sent from Chiba University to replace our faulty ones.
2.colony check
sample no . of colony 32 +++ X4 ~30 X4 7 70 0 71 4 74 5 75 7
3.min prep
sample conc. 1-15J 98 ng/uL 1-15L 87 43 169 44 172 52 114 65 214 67 148 67 126
4.transfromation
saple 59 , 72 , 68 , 69 chiba team parts ( 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9)
5.digation
K204058
Vector | Insert | ||
---|---|---|---|
1-8E | 4 | 1-8I | 10 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 | 2 | No.2 | 2 |
dH2O | 12 | dH2O | 6 |
total | 20uL | total | 20uL |
K204062
Vector | Insert | ||
---|---|---|---|
1-8E | 9 | 33 | 1 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 | 2 | No.2 | 2 |
dH2O | 7 | dH2O | 15 |
total | 20uL | total | 20uL |
K204072
Vector | Insert | ||
---|---|---|---|
1-1D | 5 | 64 | 2 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 | 2 | No.2 | 2 |
dH2O | 9 | dH2O | 12 |
total | 20uL | total | 20uL |
K204077
Vector | Insert | ||
---|---|---|---|
1-6I | 7 | 64 | 2 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 | 2 | No.2 | 2 |
dH2O | 7 | dH2O | 12 |
total | 20uL | total | 20uL |
K204078
Vector | Insert | ||
---|---|---|---|
1-2M | 6 | 1-15N | 12 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 | 2 | No.2 | 2 |
dH2O | 10 | dH2O | 4 |
total | 20uL | total | 20uL |
↓
37 degree , 2hr
gel cut & purification
ligation