"
September/13 September 2009
From 2009.igem.org
1.Min prep
Result of Nanodrop concentration check
(sample No.) (concentration)
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K113009 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 | ||
|---|---|---|---|
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0040 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 | ||
|---|---|---|---|
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0010 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 , [http://partsregistry.org/wiki/index.php?title=Part:BBa_J04450 1-1K] , [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13601 1-16I]
transformation : No.45 , No.46 , [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0060 1-4K] , [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0160 1-16C]
37 degree O/N
5.digestion and ligation
digestion with restriction enzyme
K204017
| Vector | Insert | ||
|---|---|---|---|
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0040 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 | ||
|---|---|---|---|
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_I0500 1-14N] | 10 | [http://partsregistry.org/wiki/index.php?title=Part:BBa_E0420 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 | ||
|---|---|---|---|
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_I0500 1-14N] | 10 | [http://partsregistry.org/wiki/index.php?title=Part:BBa_E0430 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
