"
August/10 August 2009
From 2009.igem.org
1.Min prep
Result of Nanodrop concentration check
(sample No.) (concentration) 1-6O 35.2 ng/uL 1-18L 41.8 ng/uL 1-10K 39.1 ng/uL
2.make competent cells(with Ca)
3.digestion and ligation
digestion with restriction enzyme
| Insert 1 | uL |
| 2-16H | 6 |
| EcoRⅠ | 1 |
| XbaⅠ | 1 |
| 10*Buffer No.2 | 2 |
| dH2O | 10 |
| TOTAL | 20 |
| Vector 1 | uL |
| 1-23L | 12 |
| EcoRⅠ | 1 |
| SpeⅠ | 1 |
| 10*Buffer No.2 | 2 |
| dH2O | 4 |
| TOTAL | 20 |
| Insert 2 | uL |
| 3-18O | 5 |
| EcoRⅠ | 1 |
| XbaⅠ | 1 |
| 10*Buffer No.2 | 2 |
| dH2O | 11 |
| TOTAL | 20 |
| Vector 2 | uL |
| 1-23L | 12 |
| EcoRⅠ | 1 |
| SpeⅠ | 1 |
| 10*Buffer No.2 | 2 |
| dH2O | 4 |
| TOTAL | 20 |
↓
failured
digestion with restriction enzyme insert : E+S vector : E+x
4.Transformation
to get new plasmid
1-23L A 1-18C A 1-20F K 1-12H K 1-6I A 2-6P A 1-15N A used '8/10 competent cell
