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24 August 2008 <|> 26 August 2008
Laboratory work
Team:Input
The following glycerol Stock was made.
BBa_J22136(insert check OK)
Picked the colony and incubate with 2mL of LB-Ampicillin Medium for 12 hours at 37 degrees.
Took 180μl from culture,and mixed with 880μlLB-Amp Medium.(OD=0.485)
Liquid culture(OD=0.495) |
0.67μl |
60%Glycerol |
0.33μl |
20%Glycerol Stock |
1.00ml |
Team:Communication
- (24/8)--->Digestion
- Double Digestion:BBa_J04500(2007)
- Double Digesiton:BBa_R0010(2007)
- Single Digestion:BBa_R0010(2007)
- BBa_R0010(2007)
Sample No. |
1 | 2 | 3 | 4 |
Sample DNA(μL) |
5 | 5 | 1 | 1 |
EcoRI(μL) |
0.5 | 0.5 | - | - |
SpeI(μL) |
0.5 | 0.5 | - | - |
Buffer 1 |
1 | 1 | - | - |
Buffer 3(μL) |
- | - | 1 | - |
BSA(μL) |
1 | 1 | - | - |
dH2O(μL) |
2 | 2 | 7.5 | - |
TOTAL(μL) |
10 | 10 | 10 | 1 |
- --->Gel Check
|
Sample No. |
1~3 | 4 |
Sample DNA(μL) |
10 | 1 |
Loading Dye(μL) |
2 | 1 |
dH2O(μL) |
- | 4 |
TOTAL(μL) |
12 | 6 |
- From right;
- Double Digestion:BBa_J04500(2007) -> OK
- Double Digesiton:BBa_R0010(2007) -> ?(low)
- Single Digestion:BBa_R0010(2007) -> ?(low)
- BBa_R0010(2007) -> OK(low)
|
--->Digestion
(Double Digestion)
- BBa_C0178
- BBa_C0170
- BBa_J04500(2007)
Sample No. |
1 | 2 | 3 |
Sample DNA(μL) |
33ng/μl×60μl | 33ng/μl×60μl | 100ng/μl×20μl |
PstI(μL) |
2 | 2 | 2 |
XbaI(μL) |
2 | 2 | - |
SpeI(μL) |
- | - | 2 |
Buffer 2(μL) |
- | - | 5 |
Buffer 3(μL) |
10 | 10 | - |
BSA(μL) |
10 | 10 | 5 |
dH2O(μL) |
16 | 16 | 16 |
TOTAL(μL) |
100 | 100 | 52 |
Team:Output
PCR
No. |
1 | 2 | 3 | 4 | 5 |
DNA tamplate(μL) |
1 | 1 | 1 | 1 | 1 |
FW primer(μL) |
5(Venus_YFP_fwd) | 5(LacZ_fwd) | 5(LacZ_fwd) | 5(rLUC_fwd) | 5(fLuc_fwd) |
VR primer(μL) |
5(VR) | 5(LacZ_rev) | 5(LacZ_rev) | 5(VR) | 5(VR) |
dNTPmix(μL) |
10 | 10 | 10 | 10 | 10 |
Thermo pol buffer(μL) |
10 | 10 | 10 | 10 | 10 |
DNA pol(VENT)(μL) |
1 | 1 | 1 | 1 | 1 |
dH2O(μL) |
68 | 68 | 68 | 68 | 68 |
TOTAL(μL) |
100 | 100 | 100 | 100 | 100 |
-->Gel Check
No. |
1 | 2 | 3 | 4 | 5 |
DNA tamplate(μL) |
1 | 1 | 1 | 1 | 1 |
loading Dye(μL) |
1 | 1 | 1 | 1 | 1 |
dH2O(μL) |
4 | 4 | 4 | 4 | 4 |
TOTAL(μL) |
6 | 6 | 6 | 6 | 6 |
Mini prep
Transformation
- BBa_E2030
- PUC19
- pGFPUV
- pGEX Venus YFP
- Membren Venus YFP
PCR
No. |
1 |
DNA tamplate(μL) |
1 |
FW primer(Venus)(μL) |
5 |
VR primer(μL) |
5 |
dNTPmix(μL) |
10 |
Thermo pol buffer(μL) |
10 |
DNA pol(VENT)(μL) |
1 |
dH2O(μL) |
68 |
TOTAL(μL) |
100 |