"
August/22 August 2009
From 2009.igem.org
1.transformed check
Checked the cell cultures transformed and plated out yesterday (8/21) for colony formation and made an approximate count of the number of colonies.
(ligation sample No.) (no. of colonies)
⑤ failure
⑦ 100~
⑧ 100~
- ⑤ try back (digestion and ligation)
- ⑦,⑧ ligation rapid check
2.Min prep
Result of Nanodrop concentration check
(sample No.) (concentration)
[http://partsregistry.org/wiki/index.php?title=Part:BBa_I732101 1-13N] 30.3 ng/uL
[http://partsregistry.org/wiki/index.php?title=Part:BBa_C0170 2-8G] 15.5
[http://partsregistry.org/wiki/index.php?title=Part:BBa_I13521 2-6O] 24.4
26.0
③-C 15.8
④-A 25.4
⑥-A 16.2
3.transfromation
ligation sample ⑨ , ⑩ , ⑪
4.digestion and ligation
K204005
| Vector | Insert | ||
|---|---|---|---|
| p(RhlR)([http://partsregistry.org/wiki/index.php?title=Part:BBa_I14017 1-18L]) | 6 | RBS+RhlR+terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_I1466 1-23J]) | 6 |
| EcoRI | 1 | EcoRI | 1 |
| XbaI | 1 | SpeI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 12 | dH2O | 12 |
| total | 20uL | total | 20uL |
K204012
| Vector | Insert | ||
|---|---|---|---|
| terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 1-23L]) | 6 | RBS+cinR(③-C) | 18 |
| EcoRI | 1 | EcoRI | 1 |
| XbaI | 1 | SpeI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 10 | dH2O | 8 |
| total | 20uL | total | 20uL |
K204013
| Vector | Insert | ||
|---|---|---|---|
| RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M]) | 10 | RhlI([http://partsregistry.org/wiki/index.php?title=Part:BBa_C0170 2-8G]) | 8 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 6 | dH2O | 8 |
| total | 20uL | total | 20uL |
K204014
| Vector | Insert | ||
|---|---|---|---|
| RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M]) | 8 | LasI([http://partsregistry.org/wiki/index.php?title=Part:BBa_C0078 1-14H]) | 4 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 8 | dH2O | 13 |
| total | 20uL | total | 20uL |
K204015
| Vector | Insert | ||
|---|---|---|---|
| terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 1-23L]) | 6 | RBS+cinI(⑥-A) | 8 |
| EcoRI | 1 | EcoRI | 1 |
| XbaI | 1 | SpeI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 10 | dH2O | 8 |
| total | 20uL | total | 20uL |
↓
37°C, overnight
