August/25 August 2009
From 2009.igem.org
1.trandformation
ligation sample No. 12 ,13 , 14 , 15 ↓ 65°C , 10min ↓ competent cell 100uL sample 10uL competent cell 100uL sample 2uL
2.Min prep
Result of Nanodrop concentration check
(sample No.) (concentration) [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0040 1-6I] 13.5 19.1 15.4 13.4 18.9 [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0051 1-4E] 15.0 15.3 14.9 [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118006 2-16N] 22.7 17.7 23.1 [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118003 2-16H] 13.8 22.7 [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118005 2-16L] 22.5 31.5
3.ligation check
sample No. 9 , 10
4.digestion
digestion with restriction enzyme
K204019
Vector | Insert | ||
---|---|---|---|
RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M]) | 8 | [http://partsregistry.org/wiki/index.php?title=Part:BBa_E2050 2-13N] | 6 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 Buffer | 2 | No.2 | 2 |
dH2O | 8 | dH2O | 10 |
total | 20uL | total | 20uL |
K204020
Vector | Insert | ||
---|---|---|---|
RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M]) | 8 | YFP([http://partsregistry.org/wiki/index.php?title=Part:BBa_E0430 1-8I]) | 6 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 Buffer | 2 | No.2 | 2 |
dH2O | 8 | dH2O | 10 |
total | 20uL | total | 20uL |
K204021
Vector | Insert | ||
---|---|---|---|
RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M]) | 8 | CFP([http://partsregistry.org/wiki/index.php?title=Part:BBa_E0420 1-8K]) | 6 |
SpeI | 1 | XbaI | 1 |
PstI | 1 | PstI | 1 |
No.2 Buffer | 2 | No.2 | 2 |
dH2O | 8 | dH2O | 10 |
total | 20uL | total | 20uL |
↓
37°C, over night