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Team:Chiba/jk/β/week 4
From 2008.igem.org
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(→10 September, 2008) |
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===10 September, 2008=== | ===10 September, 2008=== | ||
| + | -->(10/9) | ||
| + | :Colony Count | ||
| + | #LuxI | ||
| + | #LuxI with LVA | ||
| + | #Las | ||
| + | #Background(R0010) | ||
| + | #Background(C0261) | ||
| + | |||
| + | Pick colony | ||
| + | |||
| + | '''[[Team:Chiba/protocol/PCR|Colony-PCR]]''' | ||
| + | ::Colony PCR of 10 colonies from ligation plate(9/9-(2)) and 3 colonies from ligation plate(9/9-(3)) one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)). | ||
| + | |||
| + | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
| + | <tr> | ||
| + | <td width="257">DNA Template</td> | ||
| + | <td>1</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>dNTP mix</td> | ||
| + | <td>5</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>Foward Primer</td> | ||
| + | <td>0.3</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>Reverse Primer</td> | ||
| + | <td>0.3</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>taq DNA polymerase</td> | ||
| + | <td>0.5</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>Thermopol Buffer</td> | ||
| + | <td>3</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>Nuclease free water</td> | ||
| + | <td>20</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>TOTAL</td> | ||
| + | <td>30μL</td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | ?? | ||
| + | :95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃ | ||
| + | ?? | ||
===11 September, 2008=== | ===11 September, 2008=== | ||
Revision as of 08:50, 30 September 2008
>送受信班
Contents |
Week 4
>last week
7 September, 2008
-->(6/9) Digestion Test
- Sample DNA 1 : BBa_R0010(濃度未知)[http://partsregistry.org/Part:BBa_R0010]
- Sample DNA 2 : BBa_R0010(100ng/μL)[http://partsregistry.org/Part:BBa_R0010]
| Sample No | 1,2 |
| Sample DNA | 12 |
| PstⅠ | 0.5 |
| SpeⅠ | 0.5 |
| Buffer 2 | 2 |
| (×10)BSA | 2 |
| dH2O | 3 |
| TOTAL | 20 |
- --->Gel Check
Sample left right Sample DNA 5 5 Loading Dye 1 1 TOTAL 6 6
- Left side:BBa_R0010 Sample No.1
- Right side:BBa_R0010 Sample No.2
-->(8/9)
8 September, 2008
-->(7/9)
- --->Zymo Clean
- ->15μL?
9 September, 2008
-->(6/9)PCR of I9030
--->Gel Check
--->Gel Extract
--->Zymo Clean
10 September, 2008
-->(10/9)
- Colony Count
- LuxI
- LuxI with LVA
- Las
- Background(R0010)
- Background(C0261)
Pick colony
- Colony PCR of 10 colonies from ligation plate(9/9-(2)) and 3 colonies from ligation plate(9/9-(3)) one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
DNA Template 1 dNTP mix 5 Foward Primer 0.3 Reverse Primer 0.3 taq DNA polymerase 0.5 Thermopol Buffer 3 Nuclease free water 20 TOTAL 30μL ??
- 95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃
??
11 September, 2008
12 September, 2008
13 September, 2008
- Gel Check
- BBa_C0161[http://partsregistry.org/Part:BBa_Co161](2007)
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
- BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
Sample No 1~3 Sample DNA 1 Loading Dye 1 dH2O 4 TOTAL 6 - From left;
- From left;
-->Digestion
- BBa_C0161[http://partsregistry.org/Part:BBa_Co161](2007)
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
- BBa_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 >next week
>next week
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