Team:Chiba/jk/β
From 2008.igem.org
(Difference between revisions)
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| | | | ||
From left; | From left; | ||
- | ::①insert -> OK | + | ::①insert:C0178 -> OK |
- | ::②insert -> OK? | + | ::②insert:C0170 -> OK? |
- | ::③Vector -> OK | + | ::③Vector:J04500 -> OK |
|} | |} | ||
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<td>6</td></tr> | <td>6</td></tr> | ||
</table> | </table> | ||
- | |||
{|align="justify" | {|align="justify" | ||
|[[Image:Chiba-0826.JPG]] | |[[Image:Chiba-0826.JPG]] | ||
Line 615: | Line 614: | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
<td>10</td><td>10</td><td>10</td><td>10</td><td>10</td> | <td>10</td><td>10</td><td>10</td><td>10</td><td>10</td> | ||
- | |||
</tr> | </tr> | ||
</table> | </table> | ||
+ | |||
+ | |||
+ | :--->'''[[Team:Chiba/protocol/transformation|Transformation]]''' | ||
+ | ::(1)insert①C0178 | ||
+ | ::(2)insert②C0170 | ||
+ | ::(3)vector③J04500 | ||
+ | ::(4)insert①C0178+vector③J04500 | ||
+ | ::(5)insert②C0170+vector③J04500 | ||
+ | |||
Revision as of 16:06, 3 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
- Transformation
- 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
- 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
- --->Gel Check
- 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
- PCR
- Colony PCR of 5 colonies from ligation plates (27/8) and one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
From left; |
From left; |
- 28 August, 2008]
From left; |
- 29 August, 2008
- 30 August, 2008
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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