Team:Chiba/Calendar-Home/26 August 2008
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(New page: >Home | Notebook 25 August 2008 <|> 27 August 2008 ==Labora...) |
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Line 18: | Line 18: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Sample DNA</td> | + | <td>Sample DNA(μL)</td> |
<td>30</td> | <td>30</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Loading Dye</td> | + | <td>Loading Dye(μL)</td> |
<td>6</td> | <td>6</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>TOTAL</td> | + | <td>TOTAL(μL)</td> |
<td>36</td> | <td>36</td> | ||
</tr> | </tr> | ||
Line 53: | Line 53: | ||
:--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]''' | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]''' | ||
- | :::#5μl | + | |
- | + | :eluted with following amount of elution buffer | |
- | + | ::#5μl | |
+ | ::#5μl | ||
+ | ::#27μl | ||
Line 68: | Line 70: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Sample DNA</td> | + | <td>Sample DNA(μL)</td> |
<td>2</td> | <td>2</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Loading Dye</td> | + | <td>Loading Dye(μL)</td> |
<td>2</td></tr> | <td>2</td></tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>H<sub>2</sub>O(μL)</td> |
<td>8</td></tr> | <td>8</td></tr> | ||
<tr> | <tr> | ||
- | <td>TOTAL</td> | + | <td>TOTAL(μL)</td> |
<td>12</td></tr> | <td>12</td></tr> | ||
</table> | </table> | ||
Line 96: | Line 98: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Sample DNA</td> | + | <td>Sample DNA(μL)</td> |
<td>26</td> | <td>26</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>SAP</td> | + | <td>SAP(μL)</td> |
<td>1</td></tr> | <td>1</td></tr> | ||
<tr> | <tr> | ||
- | <td>SAP Buffer</td> | + | <td>SAP Buffer(μL)</td> |
<td>3</td></tr> | <td>3</td></tr> | ||
<tr> | <tr> | ||
- | <td>TOTAL</td> | + | <td>TOTAL(μL)</td> |
- | <td> | + | <td>30</td></tr> |
</table> | </table> | ||
Line 126: | Line 128: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Loading Dye</td> | + | <td>Loading Dye(μL)</td> |
<td>1</td> | <td>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>dH<sub>2</sub>O</td> | + | <td>dH<sub>2</sub>O(μL)</td> |
<td>4</td> | <td>4</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>TOTAL</td> | + | <td>TOTAL(μL)</td> |
<td>6</td></tr> | <td>6</td></tr> | ||
</table> | </table> | ||
Line 148: | Line 150: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>insert(1)[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](μL)</td> |
<td>1</td><td>-</td><td>-</td><td>1</td><td>-</td> | <td>1</td><td>-</td><td>-</td><td>1</td><td>-</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>insert(2)[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](μL)</td> |
<td>-</td><td>1</td><td>-</td><td>-</td><td>1</td> | <td>-</td><td>1</td><td>-</td><td>-</td><td>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>vector(3)[http://partsregistry.org/Part:BBa_J04500 BBa_J04500](μL)</td> |
<td>-</td><td>-</td><td>2</td><td>2</td><td>2</td> | <td>-</td><td>-</td><td>2</td><td>2</td><td>2</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>ligase</td> | + | <td>ligase(μL)</td> |
<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td> | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Buffer</td> | + | <td>Buffer(μL)</td> |
<td>2</td><td>2</td><td>2</td><td>2</td><td>2</td> | <td>2</td><td>2</td><td>2</td><td>2</td><td>2</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>dH<sub>2</sub>O</td> | + | <td>dH<sub>2</sub>O(μL)</td> |
<td>6</td><td>6</td><td>5</td><td>4</td><td>4</td> | <td>6</td><td>6</td><td>5</td><td>4</td><td>4</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>TOTAL</td> | + | <td>TOTAL(μL)</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> | ||
Line 179: | Line 181: | ||
:--->'''[[Team:Chiba/protocol/transformation|Transformation]]''' | :--->'''[[Team:Chiba/protocol/transformation|Transformation]]''' | ||
- | ::# | + | ::#insert(1)[http://partsregistry.org/Part:BBa_C0178 BBa_C0178] |
- | ::# | + | ::#insert(2)[http://partsregistry.org/Part:BBa_C0170 BBa_C0170] |
- | ::# | + | ::#vector(3)[http://partsregistry.org/Part:BBa_J04500 BBa_J04500] |
- | ::# | + | ::#insert(1)[http://partsregistry.org/Part:BBa_C0178 BBa_C0178]+vector(3)[http://partsregistry.org/Part:BBa_J04500 BBa_J04500] |
- | ::# | + | ::#insert(2)[http://partsregistry.org/Part:BBa_C0170 BBa_C0170]+vector(3)[http://partsregistry.org/Part:BBa_J04500 BBa_J04500] |
- | + | ||
- | + | ||
===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><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> | + | <td>6μl</td><td>6μl</td><td>6μl</td> |
</tr> | </tr> | ||
</table> | </table> | ||
[[Team:Chiba/protocol/digestion|Digestion]] | [[Team:Chiba/protocol/digestion|Digestion]] | ||
- | *vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] | + | *vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](1) |
<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> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 228: | Line 228: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>SpeI</td> |
<td>2</td> | <td>2</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>PstI</td> |
<td>2</td> | <td>2</td> | ||
</tr> | </tr> | ||
Line 245: | Line 245: | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>130μl</td> |
</tr> | </tr> | ||
</table> | </table> | ||
- | *insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008] | + | *insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](1) |
+ | :[http://partsregistry.org/Part:BBa_E2030 BBa_E2030](2) | ||
<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> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 261: | Line 262: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>XbaI</td> |
<td>1</td><td>1</td> | <td>1</td><td>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>PstI</td> |
<td>1</td><td>1</td> | <td>1</td><td>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>DpnI</td> |
<td>1</td><td>1</td> | <td>1</td><td>1</td> | ||
</tr> | </tr> | ||
Line 282: | Line 283: | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>40μl</td><td>40μl</td> |
</tr> | </tr> | ||
</table> | </table> | ||
- | <br>--> | + | <br>-->37°C 3hour |
<br>-->SAP | <br>-->SAP | ||
- | <br>--> | + | <br>-->37°C 30min |
- | <br>--> | + | <br>-->65°C 15min |
<br>-->[[Team:Chiba/protocol/DNA_Purification/zymocleam|Zymo Clean]] | <br>-->[[Team:Chiba/protocol/DNA_Purification/zymocleam|Zymo Clean]] | ||
[[Team:Chiba/protocol/ligation/ligation|Ligation]] | [[Team:Chiba/protocol/ligation/ligation|Ligation]] | ||
- | *vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008] | + | *vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](1) |
- | *vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_E2030 BBa_E2030] | + | *vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_E2030 BBa_E2030](2) |
- | *Negative control:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] | + | *Negative control:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](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> | ||
Line 324: | Line 325: | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>20μl</td><td>20μl</td><td>20μl</td> |
</tr> | </tr> | ||
</table> | </table> | ||
Line 332: | Line 333: | ||
-->[[Team:Chiba/protocol/transformation|Transformation]](XL10G) | -->[[Team:Chiba/protocol/transformation|Transformation]](XL10G) | ||
- | --> | + | -->37°C over night |
-->colony PCR | -->colony PCR | ||
-->[[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]] | -->[[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]] |
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
|
|
|
--->Gel Extract
- --->Zymo Clean
- eluted with following amount of elution buffer
- 5μl
- 5μl
- 27μl
- --->Gel Check
|
- --->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