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Team:Chiba/Calendar-Home/26 August 2008
From 2008.igem.org
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===Team:Output=== | ===Team:Output=== | ||
[[Team:Chiba/protocol/gelcheck|Gel Check]] | [[Team:Chiba/protocol/gelcheck|Gel Check]] | ||
| - | *[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] | + | *[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](1) |
| - | *[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] | + | *[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](2) |
| - | *[http://partsregistry.org/Part:BBa_J63001 BBa_J63001] | + | *[http://partsregistry.org/Part:BBa_J63001 BBa_J63001](3) |
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
<td width="257">Sample No.</td> | <td width="257">Sample No.</td> | ||
| - | <td> | + | <td>1</td><td>2</td><td>3</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>DNA tamplate</td> | <td>DNA tamplate</td> | ||
| - | + | <td>1</td><td>1</td><td>1</td> | |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Loading Dye</td> | <td>Loading Dye</td> | ||
| - | + | <td>1</td><td>1</td><td>1</td> | |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>dH<sub>2</sub>O</td> | <td>dH<sub>2</sub>O</td> | ||
| - | + | <td>4</td><td>4</td><td>4</td> | |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
| - | + | <td>6μl</td><td>6μl</td><td>6μl</td> | |
</tr> | </tr> | ||
</table> | </table> | ||
Latest revision as of 01:04, 30 October 2008
25 August 2008 <|> 27 August 2008
Contents |
Laboratory work
Team:Input
no work
Team:Communication
- (8/25)--->Gel Check
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--->Gel Extract
- --->Zymo Clean
- eluted with following amount of elution buffer
- 5μl
- 5μl
- 27μl
- --->Gel Check
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- --->SAP
Sample No. 3 Sample DNA(μL) 26 SAP(μL) 1 SAP Buffer(μL) 3 TOTAL(μL) 30
- --->Zymo Clean
- Sample No.3 -->9μl
- --->Gel Check
Sample No.3 1 Loading Dye(μL) 1 dH2O(μL) 4 TOTAL(μL) 6 - Sample No.3 -> OK ･･･30ng?
- --->Ligation
Sample No. (1) (2) (3) (4) (5) insert(1)BBa_C0178(μL) 1 - - 1 - insert(2)BBa_C0170(μL) - 1 - - 1 vector(3)BBa_J04500(μL) - - 2 2 2 ligase(μL) 1 1 1 1 1 Buffer(μL) 2 2 2 2 2 dH2O(μL) 6 6 5 4 4 TOTAL(μL) 10 10 10 10 10
- --->Transformation
- insert(1)BBa_C0178
- insert(2)BBa_C0170
- vector(3)BBa_J04500
- insert(1)BBa_C0178+vector(3)BBa_J04500
- insert(2)BBa_C0170+vector(3)BBa_J04500
Team:Output
- BBa_F2620(1)
- BBa_F2620(2)
- BBa_J63001(3)
Sample No. 1 2 3 DNA tamplate 1 1 1 Loading Dye 1 1 1 dH2O 4 4 4 TOTAL 6μl 6μl 6μl - vector:BBa_F2620(1)
Sample No. 1 DNA tamplate 100 SpeI 2 PstI 2 BSA(x10) 13 Buffer2 13 TOTAL 130μl - insert:BBa_J52008(1)
- BBa_E2030(2)
Sample No. 1 2 DNA tamplate 30 30 XbaI 1 1 PstI 1 1 DpnI 1 1 Buffer3 4 4 BSA(x10) 4 4 TOTAL 40μl 40μl
-->37°C 3hour
-->SAP
-->37°C 30min
-->65°C 15min
-->Zymo Clean- vector:BBa_F2620 insert:BBa_J52008(1)
- vector:BBa_F2620 insert:BBa_E2030(2)
- Negative control:BBa_F2620(3)
Sample No. 1 2 3 vector 5 5 5 inser 10 7.5 0 Ligase Buffer 4 4 4 Ligase 1 1 1 dH2O 0 2.5 10 TOTAL 20μl 20μl 20μl -->RT 2hour
-->Transformation(XL10G)
-->37°C over night
-->colony PCR
-->Mini prep
- --->Zymo Clean
