August/7 October 2009
From 2009.igem.org
1. colony check
sample no. no. of colony 11 +++ 20 +++ 21 +++ 2-6O ++ 1-15J + 1-15L +
2.transformation
sample no. 67 , 68 , 69 , 52 , 11 , 20 , 21 , 2-6O , 1-15J , 1-15L
3.digation
K204070
Vector | Insert | ||
---|---|---|---|
33 | 2 | 50 | 5 |
EcoRI | 1 | EcoRI | 1 |
XbaI | 1 | SpeI | 1 |
No.2 | 2 | No.2 | 2 |
dH2O | 12 | dH2O | 9 |
total | 20uL | total | 20uL |
K204071
Vector | Insert | ||
---|---|---|---|
1-8K | 2 | 51 | 4 |
EcoRI | 1 | EcoRI | 1 |
XbaI | 1 | SpeI | 1 |
No.2 | 2 | No.2 | 2 |
dH2O | 12 | dH2O | 10 |
total | 20uL | total | 20uL |
K204072
Vector | Insert | ||
---|---|---|---|
2-8E | 2 | 31 | 6 |
EcoRI | 1 | EcoRI | 1 |
XbaI | 1 | SpeI | 1 |
No.2 | 2 | No.2 | 2 |
dH2O | 12 | dH2O | 12 |
total | 20uL | total | 20uL |
K204073
Vector | Insert | ||
---|---|---|---|
1-8I | 6 | 32 | 6 |
EcoRI | 1 | EcoRI | 1 |
XbaI | 1 | SpeI | 1 |
No.2 | 2 | No.2 | 2 |
dH2O | 8 | dH2O | 8 |
total | 20uL | total | 20uL |
K204074
Vector | Insert | ||
---|---|---|---|
1-1K | 10 | 1-12M | 2 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 | 2 | No.2 | 2 |
dH2O | 4 | dH2O | 12 |
total | 20uL | total | 20uL |
K204075
Vector | Insert | ||
---|---|---|---|
1-1K | 10 | 1-8K | 2 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 | 2 | No.2 | 2 |
dH2O | 4/td> | dH2O | 12 |
total | 20uL | total | 20uL |
↓
37 degree . O/N
4. min prep
sample conc. X4 257 ng/uL X4 266 X4 171 X5 169 X6 293 X7 294 X8 151 59 306 61 285 64 252 65 278
5.digation
X4
Vector | Insert | ||
---|---|---|---|
32 | 16 | 1-12M | 4 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 | 2 | No.2 | 2 |
dH2O | 2 | dH2O | 12 |
total | 20uL | total | 20uL |
↓
37 degree , 2hr
gel cut