September/13 September 2009
From 2009.igem.org
1.Min prep
Result of Nanodrop concentration check
(sample No.) (concentration) 3-1M 27.6 ng/uL No. 30 162.0 ng/uL No. 36 67.6 ng/uL
2.digestion and ligation
digestion with restriction enzyme
K204047
Vector | Insert | ||
---|---|---|---|
1-6I | 10 | No. 24 | 6 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 Buffer | 2 | No.2 | 2 |
dH2O | 6 | dH2O | 10 |
total | 20uL | total | 20uL |
K204048
Vector | Insert | ||
---|---|---|---|
1-1D | 5 | No. 33 | 5 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 Buffer | 2 | No.2 | 2 |
dH2O | 11 | dH2O | 11 |
total | 20uL | total | 20uL |
3.ligation check
UP : ligation sample No.47 No.48
DOWN : for ligation check 14A 14B 27 28 29 30 35B 36 39
No. 47 , 48 gel purification & litaion 16 degree O/N
ligation check : failure ???
4.transfromation
for DNA modification : No.41 , No.42 , 1-1K , 1-16I
transformation : No.45 , No.46 , 1-4K , 1-16C
37 degree O/N
5.digestion and ligation
digestion with restriction enzyme
K204017
Vector | Insert | ||
---|---|---|---|
1-6I | 4 | No.9 | 8 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 Buffer | 2 | No.2 | 2 |
dH2O | 12 | dH2O | 18 |
total | 20uL | total | 20uL |
K204037
Vector | Insert | ||
---|---|---|---|
1-14N | 10 | 1-8K | 2 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 Buffer | 2 | No.2 | 2 |
dH2O | 6 | dH2O | 14 |
total | 20uL | total | 20uL |
K204038
Vector | Insert | ||
---|---|---|---|
1-14N | 10 | 1-8I | 2 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 Buffer | 2 | No.2 | 2 |
dH2O | 6 | dH2O | 14 |
total | 20uL | total | 20uL |
37 degree O/N