Team:Chiba/jk/β/week 1
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Contents |
Week 1
17 August, 2008
- Transformation
- competent cells : XL10G
18 August, 2008
DNA Template | 1 |
dNTP mix | 10 |
Foward Primer | 5 |
Reverse Primer | 5 |
DNA polymerase | 1 |
Thermopol Buffer | 10 |
dH2O | 68 |
TOTAL | 100μL |
- 95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃
- --->Gel Check
competent cells : XL10G
- --->(20/8)Mini prep
19 August, 2008
DNA Template | 1 |
dNTP mix | 10 |
Foward Primer | 5 |
Reverse Primer | 5 |
DNA polymerase VENT | 1 |
Thermopol Buffer | 5 |
dH2O | 28 |
TOTAL | 50μL |
- 95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃
- --->Gel Check
20 August, 2008
- (18/8)--->Mini prep
- --->Gel Check
--->Digestion test
Sample No | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
BBa_C0170(2006) [68] | 3 | - | - | - | 3 | - | - | - |
BBa_C0170(2007) [69] | - | 3 | - | - | - | 3 | - | - |
BBa_C0178(2006) [70] | - | - | 3 | - | - | - | 3 | - |
BBa_C0178(2007) [71] | - | - | - | 3 | - | - | - | 3 |
EcoRⅠ | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 |
PstⅠ | - | - | - | - | 0.1 | 0.1 | 0.1 | 0.1 |
Buffer EcoRⅠ | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
BSA | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
dH2O | 4.9 | 4.9 | 4.9 | 4.9 | 4.8 | 4.8 | 4.8 | 4.8 |
TOTAL | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
- --->Gel Check
|
21 August, 2008
- (20/8)--->PCR
DNA Template | 1 |
dNTP mix | 10 |
Foward Primer | 5 |
Reverse Primer | 5 |
DNA polymerase TAQ | 1 |
Thermopol Buffer | 5 |
dH2O | 28 |
TOTAL | 50μL |
- 95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃
- --->Gel Check
- Transformation
- competent cells : XL10G
--->(23/8)Digestion
22 August, 2008
- Transformation
- competent cells : XL10G
- (Locate & 2 Punch ~iyama ver.~)
--->(24/8)Mini prep
23 August, 2008
- (21/8)--->Mini prep
- --->Digestion test
Sample No 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 BBa_I9030(2007) [117] 3 - - - - - - 3 - - - - - - 3 - - - - - - BBa_I9030(2006) [118] - 3 - - - - - - 3 - - - - - - 3 - - - - - BBa_S03154(2007) [119] - - 3 - - - - - - 3 - - - - - - 3 - - - - BBa_S03154(2006) [120] - - - 3 - - - - - - 3 - - - - - - 3 - - - BBa_I9026(2007) [121] - - - - 3 - - - - - - 3 - - - - - - 3 - - BBa_I9026(2007) [122] - - - - - 3 - - - - - - 3 - - - - - - 3 - BBa_J04500(2007) [123] - - - - - - 1 - - - - - - 3 - - - - - - 3 EcoRⅠ - - - - - - - 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 PstⅠ - - - - - - - - - - - - - - 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Buffer EcoRⅠ - - - - - - - 0.9 0.9 0.9 0.9 0.9 0.9 0.9 0.8 0.8 0.8 0.8 0.8 0.8 0.8 BSA - - - - - - - 1 1 1 1 1 1 1 1 1 1 1 1 1 1 dH2O - - - - - - - 5 5 5 5 5 5 5 5 5 5 5 5 5 5 TOTAL 3 3 3 3 3 3 1 10 10 10 10 10 10 10 10 10 10 10 10 10 10
- --->Gel Check
Sample No 8~21 Sample DNA 10 Loading Dye 2 TOTAL 12 - From left;
- (Single Digestion) Sanple No. 8~14
- Digestion test
- BBa_J04500(2007:old-sample)[145]
Sample No 1 2 3 Sample DNA 3 3 3 EcoRⅠ - 0.1 0.1 SpeⅠ - - 0.1 EcoRⅠ Buffer - 0.9 0.8 BSA - 1 1 dH2O - 5 5 TOTAL 3 10 10
--->Gel Check>next week
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