Team:Chiba/jk/β
From 2008.igem.org
(Difference between revisions)
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<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>100μL</td> |
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<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>12μL</td> |
</tr> | </tr> | ||
</table> | </table> | ||
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<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>12μL</td> |
</tr> | </tr> | ||
</table> | </table> | ||
{|align="justify" | {|align="justify" | ||
- | |[[Image:Chiba- | + | |[[Image:Chiba-0819.JPG]] |
| | | | ||
::*BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] ->None | ::*BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] ->None |
Revision as of 14:10, 11 September 2008
β班の実験ノート
Contents |
Week 1
- 17 August, 2008
- Transformation
- BBa_R0040(2008) [http://partsregistry.org/Part:BBa_R0040] -> No colonies on plates
- BBa_C0070(2008) [http://partsregistry.org/Part:BBa_C0070] -> No colonies on plates
- BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates
- BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] -> No colonies on plates
- BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] -> No colonies on plates
- BBa_C0079(2008) [http://partsregistry.org/Part:BBa_C0079] -> No colonies on plates
- 18 August, 2008
- PCR
- BBa_C0070(2008) [http://partsregistry.org/Part:BBa_C0070]
- BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076]
- BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077]
- BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078]
- BBa_C0079(2008) [http://partsregistry.org/Part:BBa_C0079]
- BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]・・・control
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
Sample DNA | 3 |
Loading Dye | 2 |
dH2O | 7 |
TOTAL | 12μL |
compitent cell : XL10G
- BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
- BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070]
- BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076]
- BBa_C0076(2006) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates(19/8)
- BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
- BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]
- BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
- BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
- BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
- BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]
- --->(20/8)Mini prep
- 19 August, 2008
- PCR
- BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
- BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
- BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
- BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
- BBa_S3154(2007) [http://partsregistry.org/Part:BBa_S3154]
- BBa_S3154(2006) [http://partsregistry.org/Part:BBa_S3154]
- BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]・・・control
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
Sample DNA | 3 |
Loading Dye | 2 |
dH2O | 7 |
TOTAL | 12μL |
- 20 August, 2008
- (18/8)--->Mini prep
- BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
- BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070]
- BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076]
- BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
- BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]
- BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
- BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
- BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
- BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]
- --->Gel Check
Sample No | ① | ② | ③ | ④ |
Sample DNA | 3 | 3 | 3 | 3 |
Loading Dye | 2 | 2 | 2 | 2 |
dH2O | 7 | 7 | 7 | 7 |
TOTAL | 12 | 12 | 12 | 12 |
|
- --->Digestion
- BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
- BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070]
- BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
- BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]
- 21 August, 2008
- (20/8)--->PCR
- BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
- BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070]
- BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076]
- BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
- BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]
- --->Gel Check
- Transformation
- BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
- BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
- BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
- BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
- BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
- BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
- 22 August, 2008
- Transformation
- BBa_J13002(2007) [http://partsregistry.org/Part:BBa_J13002]
- BBa_J13002(2006) [http://partsregistry.org/Part:BBa_J13002]
- BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]
- BBa_J04500(2006) [http://partsregistry.org/Part:BBa_J04500]
- (Locate & 2 Punch ~iyama ver.~)
- BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates
- BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] -> No colonies on plates
- BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] -> No colonies on plates
- 23 August, 2008
- --->Mini prep
- BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
- BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
- BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
- BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
- BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
- BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
- --->Digestion
- BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
- BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
- BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
- BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
- BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
- BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
- BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]
- --->Gel Check</table>
Week 2
- 24 August, 2008
- (22/8)--->Mini prep
- BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
- BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
- --->Digestion
- BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
- BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
- --->Gel Check
Sample No | ① | ② | ③ | ④ | ⑤ | ⑥ | ⑦ | ⑧ |
Sample DNA | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
Loading Dye | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
dH2O | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 |
TOTAL | 6 | 6 | 6 | 6 | 6 | 6 | 6 | 6 |
Sample No | ⑤ | ⑥ | ⑦ | ⑧ |
Sample DNA | 9 | 9 | 9 | 9 |
Loading Dye | 2 | 2 | 2 | 2 |
dH2O | 1 | 1 | 1 | 1 |
TOTAL | 12 | 12 | 12 | 12 |
- 25 August, 2008
- (24/8)--->Digestion
- ①Double Digestion:BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
- ②Double Digesiton:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
- ③Single Digestion:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
- ④BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
Sample No | ① | ② | ③ | ④ |
Sample DNA | 5 | 5 | 1 | 1 |
EcoRⅠ | 0.5 | o.5 | - | - |
SpeⅠ | 0.5 | 0.5 | - | - |
Buffer 1 | 1 | 1 | - | - |
Buffer 3 | - | - | 1 | - |
BSA | 1 | 1 | - | - |
dH2O | 2 | 2 | 7.5 | - |
TOTAL | 10 | 10 | 10 | 1 |
- --->Gel Check
Sample No | ① | ② | ③ | ④ |
Sample DNA | 10 | 10 | 10 | 10 |
Loading Dye | 2 | 2 | 2 | 1 |
dH2O | - | - | - | 4 |
TOTAL | 12 | 12 | 12 | 6 |
--->Digestion
- ①BBa_C0178[http://partsregistry.org/Part:BBa_C0178]
- ②BBa_C0170[http://partsregistry.org/Part:BBa_C0170]
- ③BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
Sample No | ① | ② | ③ |
Sample DNA | 33ng/μl×60μl | 33ng/μl×60μl | 100ng/μl×20μl |
PstⅠ | 2 | 2 | 2 |
XbaⅠ | 2 | 2 | - | SpeⅠ | - | - | 2 |
Buffer 2 | - | - | 5 |
Buffer 3 | 10 | 10 | - |
BSA | 10 | 10 | 5 |
dH2O | 16 | 16 | 16 |
TOTAL | 100 | 100 | 52 |
- --->Gel Check
Sample No | ① | ② | ③ |
Sample DNA | 30 | 30 | 30 |
Loading Dye | 6 | 6 | 6 |
TOTAL | 36 | 36 | 36 |
|
|
|
- 26 August, 2008
(8/25)--->Gel Extract
- --->Zymo Clean
- ①5μl
- ②5μl
- ③27μl
- --->Gel Check
Sample No | ① | ② | ③ |
Sample DNA | 2 | 2 | 2 |
Loading Dye | 2 | 2 | 2 |
H2O | 8 | 8 | 8 |
TOTAL | 12 | 12 | 12 |
From left;
|
- --->Dephosphorylation
- ③
- --->Zymo Clean
- ③->9μl
- --->Gel Check
Sample DNA ③ | 1 |
Loading Dye | 1 |
dH2O | 4 |
TOTAL | 6 |
③ -> OK ・・・30ng? |
- --->Ligation
Sample No | (1) | (2) | (3) | (4) | (5) |
insert①C0178 | 1 | - | - | 1 | - |
insert②C0170 | - | 1 | - | - | 1 |
vector③J04500 | - | - | 2 | 2 | 2 | ligase | 1 | 1 | 1 | 1 | 1 |
Buffer | 2 | 2 | 2 | 2 | 2 |
dH2O | 6 | 6 | 5 | 4 | 4 |
TOTAL | 10 | 10 | 10 | 10 | 10 |
- --->Transformation
- (1)insert①C0178
- (2)insert②C0170
- (3)vector③J04500
- (4)insert①C0178+vector③J04500
- (5)insert②C0170+vector③J04500
- 27 August, 2008
- Sample DNA : BBa_R0010[http://partsregistry.org/Part:BBa_R0010]
Sample No | ① | ② | ③ |
Sample DNA | 1 | 1 | 1 |
PstⅠ | - | - | 0.5 |
EcoRⅠ | - | 0.5 | 0.5 |
Buffer 3 | - | 1 | 1 |
BSA | - | - | 1 |
dH2O | - | 7.5 | 2 |
TOTAL | 1 | 10 | 10 |
- --->Gel Check
Sample No | ① | ② | ③ |
Sample DNA | 1 | 5 | 5 |
Loading Dye | 1 | 1 | 1 |
H2O | 4 | - | - |
TOTAL | 6 | 6 | 6 |
From left;
|
- (26/8)--->PCR
- Colony PCR of 5 colonies from ligation plates (26/8-(4),(5)) and one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
Sample No | (4)-1~5 | (5)-1~5 | (F2620) |
insert①C0178 | 1 | - | - |
insert②C0170 | - | 1 | - |
vector③J04500 | - | - | 2 | ligase | 1 | 1 | 1 |
Buffer | 2 | 2 | 2 |
dH2O | 6 | 6 | 5 |
TOTAL | 10 | 10 | 10 |
- --->Gel Check
Sample DNA | 10 |
Loading Dye | 2 |
TOTAL | 12 |
From left;
|
- --->(28/8)Mini prepDigestion Test
Transformation
comptent cell : XL10G
- BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
- BBa_C0062(2006) [http://partsregistry.org/Part:BBa_C0062] -> no colony on plates
- BBa_C0179(2007) [http://partsregistry.org/Part:BBa_C0179]
- BBa_C0179(2006) [http://partsregistry.org/Part:BBa_C0179]
- BBa_S03156(2007) [http://partsregistry.org/Part:BBa_S03156]
- BBa_S03156(2006) [http://partsregistry.org/Part:BBa_S03156]
- BBa_S03158(2007) [http://partsregistry.org/Part:BBa_S03158]
- BBa_S03158(2006) [http://partsregistry.org/Part:BBa_S03158]
- BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160]
- BBa_S0316(2006) [http://partsregistry.org/Part:BBa_S03160]
- --->Mini prep
- --->(29/8)Gel Check
- 28 August, 2008]
(27/8)--->Mini prep
- ①insert:C0170 + vector:J04500
- ②insert:C0178 + vector:J04500
- --->Digestion Test
- --->(29/8)Gel Check
Sample No | ①② | Single Digesiton:①② | Double Digesiton:①② |
Sample DNA | 1 | 10 | 10 |
Loading Dye | 1 | 2 | 2 |
H2O | 4 | - | - |
TOTAL | 6 | 12 | 12 |
From left;
|
- 29 August, 2008
- --->Digestion Test
- ①BBa_R0010(2007) [http://partsregistry.org/Part:BBa_R0010]
- ②BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
- ③BBa_C0179(2007) [http://partsregistry.org/Part:BBa_C0179]
- ④BBa_C0179(2006) [http://partsregistry.org/Part:BBa_C0179]
- ⑤BBa_S03156(2007) [http://partsregistry.org/Part:BBa_S03156]
- ⑥BBa_S03156(2006) [http://partsregistry.org/Part:BBa_S03156]
- ⑦BBa_S03158(2007) [http://partsregistry.org/Part:BBa_S03158]
- ⑧BBa_S03158(2006) [http://partsregistry.org/Part:BBa_S03158]
- ⑨BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160]
- ⑩BBa_S03160(2006) [http://partsregistry.org/Part:BBa_S03160]
Sample No | S①~⑩ | w① | w②~⑩ |
Sample DNA | 1 | 4 | 2 |
PstⅠ | - | 0.075 | 0.1 |
EcoRⅠ | 0.1 | 0.075 | 0.1 |
Buffer 3 | 1 | 0.75 | 1 |
BSA | - | 0.75 | 1 |
dH2O | 7.9 | 4.35 | 5.8 |
TOTAL | 10 | 10 | 10 |
- --->Gel Check
Sample DNA | 10 |
Loading Dye | 2 |
TOTAL | 12 |
From left; |
From left; |
(27/8)--->
R0010 |
- ①BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
- ②BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
- ③BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
- ④BBa_R0010(2006) [http://partsregistry.org/Part:BBa_R0010]
Sample No | S①~③ | ④ |
Sample DNA | 100ng/μL×20 | 200ng/μL×30 |
PstⅠ | 2 | 2 |
XbaⅠ | 2 | - |
SpeⅠ | - | 2 |
Buffer 2 | - | 5 |
Buffer 3 | 5 | - |
BSA | 5 | 5 |
dH2O | 16 | 6 |
TOTAL | 50 | 50 |
- --->Gel Check
Sample DNA | 10 | 3 |
Loading Dye | 2 | 2 |
dH2O | - | 7 |
TOTAL | 12 |
From left; |
From left; |
From left; |
From left; |
)--->Gel Extract
- --->Zymo Clean
- =>1.5μL/2colum
- --->SAP
④
- --->Zymo Clean
- ---> miss!
- 30 August, 2008
Transformation comptent cell : XL10G
- BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
- BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
- BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
- BBa_S03156(2007) [http://partsregistry.org/Part:BBa_S03156]
- BBa_S03158(2007) [http://partsregistry.org/Part:BBa_S03158]
- BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160]
- BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
- BBa_C0179(2007) [http://partsregistry.org/Part:BBa_C0179]
- --->(31/8)Mini prep
(29/8)--->Mini prep
- ①insert:C0170 + vector:J04500
- ②insert:C0178 + vector:J04500
- --->Digestion Test
- --->Gel Check
From left; |
From left; |
From left; |
Week 3
- 31 August, 2008
- 1 September, 2008
- 2 September, 2008
- 3 September, 2008
- 4 September, 2008
- 5 September, 2008
- 6 September, 2008
Week 4
- 7 September, 2008
- 8 September, 2008
- 9 September, 2008
- 10 September, 2008
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