Team:Chiba/Calendar-Home/25 September 2008
From 2008.igem.org
24 September 2008 <|> 26 September 2008
Laboratory work
Team:Communication
Team:Output
colony PCR
Vector:BBa_R0062 Insert:BBa_E0240
Vector:BBa_R0062
Sample No | each |
culture | 1 |
Fwd primer | 1.5 |
Rev primer | 1.5 |
Taq(DNA pol) | 0.2 |
dNTPmix | 3 |
thermo pol Buffer | 3 |
dH2O | 19.8 |
TOTAL | 30 |
->95℃ 5min ->(95℃ 1min 50℃ 30sec 72℃ 1min)x25 ->72℃ 10min
Digestion Test
BBa_I6206
BBa_I6101
BBa_I6042
BBa_I0462
Sample No | each |
DNA | 2 |
EcoRⅠ | 0.2 |
EcoRⅠ Buffer | 1 |
dH2O | 7 |
TOTAL | 30 |
->30min 37℃
Gel cheack
BBa_I6206(digestion product)
BBa_I6101(digestion product)
BBa_I6042(digestion product)
BBa_I0462(digestion product)
BBa_I0642(natural)
Sample No | each(digestion) | I0462(natural) |
DNA | 10 | 1 |
Loading Dye | 2 | 1 |
dH2O | - | 4 |
TOTAL | 12 | 6 |
Digestion
BBa_I0462
DNA | 25 |
XbaⅠ | 1 |
PstⅠ | 1 |
Buffer 3 | 4 |
BSA | 4 |
dH2O | 5 |
TOTAL | 40 |
->3hour 37℃
->Zymo ->10μl
Ligation
Vector:BBa_I7100
Insert:BBa_R0062
Insert:BBa_I0462
I7100 | 1.3 | 1.3 |
R0062 | 2 | - |
I0462 | 3.3 | - |
Ligase | 1 | 1 |
Ligase Buffer | 2 | 2 |
dH2O | 0.4 | 5 |
TOTAL | 10 | 10 |
->3hour 37℃
->Transformation