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Team:Chiba/Calendar-Home/19 September 2008
From 2008.igem.org
(Difference between revisions)
| Line 13: | Line 13: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>Sample DNA( | + | <td>Sample DNA(μL)</td> |
<td>30</td><td>30</td><td>90</td><td>?</td> | <td>30</td><td>30</td><td>90</td><td>?</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>Loading Dye( | + | <td>Loading Dye(μL)</td> |
<td>6</td><td>6</td><td>18</td><td>?</td> | <td>6</td><td>6</td><td>18</td><td>?</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>TOTAL( | + | <td>TOTAL(μL)</td> |
<td>36</td><td>36</td><td>108</td><td>24</td> | <td>36</td><td>36</td><td>108</td><td>24</td> | ||
</tr> | </tr> | ||
| Line 27: | Line 27: | ||
--->'''[[Team:Chiba/protocol/ligation/gelex|Gel Extract]]'''<br> | --->'''[[Team:Chiba/protocol/ligation/gelex|Gel Extract]]'''<br> | ||
| - | ( | + | (3 wells for 1 sample)<br> |
| - | *note:ADB buffer添加後, | + | *note:ADB buffer添加後,left at 37°C,for 6 hours |
:--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]''' | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]''' | ||
*elution | *elution | ||
| - | **J0450--> | + | **J0450-->27μL |
| - | **C0161--> | + | **C0161-->11μL |
--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | --->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | ||
| Line 43: | Line 43: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>C0161( | + | <td>C0161(μL)</td> |
<td>3</td><td>3</td><td>-</td> | <td>3</td><td>3</td><td>-</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>J04500( | + | <td>J04500(μL)</td> |
<td>4.5</td><td>-</td><td>4.5</td> | <td>4.5</td><td>-</td><td>4.5</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>ligase( | + | <td>ligase(μL)</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(invitrogen)( | + | <td>Buffer(invitrogen)(μL)</td> |
<td>3</td><td>2</td><td>2</td> | <td>3</td><td>2</td><td>2</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>NFW | + | <td>NFW(μL)</td> |
<td>3.5</td><td>4</td><td>2.5</td> | <td>3.5</td><td>4</td><td>2.5</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>TOTAL( | + | <td>TOTAL(μL)</td> |
<td>15</td><td>10</td><td>10</td> | <td>15</td><td>10</td><td>10</td> | ||
</tr> | </tr> | ||
| Line 71: | Line 71: | ||
*note | *note | ||
**competent cell:XL10GOLD | **competent cell:XL10GOLD | ||
| - | **DNA: | + | **DNA:1μL,SOC:120μL |
--->(20/9)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]] | --->(20/9)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]] | ||
| Line 81: | Line 81: | ||
#temparature | #temparature | ||
| - | + | 30°C and Room Temparature | |
===Team:Output=== | ===Team:Output=== | ||
<br>colony PCR | <br>colony PCR | ||
| - | <br>ptet+LuxR( | + | <br>ptet+LuxR(×16) |
| - | <br>ptet+LasR( | + | <br>ptet+LasR(×7) |
| - | <br>ptet+RhlR( | + | <br>ptet+RhlR(×5) |
| - | <br>ptet+cinR( | + | <br>ptet+cinR(×5) |
Revision as of 19:53, 29 October 2008
18 September 2008 <|> 20 September 2008
Laboratory work
Team:Communication
-->(?/9) Gel Check
| Sample Name | J04500 | R0010 | C0161 | Cin |
| Sample DNA(μL) | 30 | 30 | 90 | ? |
| Loading Dye(μL) | 6 | 6 | 18 | ? |
| TOTAL(μL) | 36 | 36 | 108 | 24 |
--->Gel Extract
(3 wells for 1 sample)
- note:ADB buffer添加後,left at 37°C,for 6 hours
- --->Zymo Clean
- elution
- J0450-->27μL
- C0161-->11μL
--->Gel Check
- --->Ligation
| Sample Name | insert+vector | insert | vector |
| C0161(μL) | 3 | 3 | - |
| J04500(μL) | 4.5 | - | 4.5 |
| ligase(μL) | 1 | 1 | 1 |
| Buffer(invitrogen)(μL) | 3 | 2 | 2 |
| NFW(μL) | 3.5 | 4 | 2.5 |
| TOTAL(μL) | 15 | 10 | 10 |
- --->Transformed above ligation onto a LB-amp plate.
- note
- competent cell:XL10GOLD
- DNA:1μL,SOC:120μL
--->(20/9)Mini prep
- time-delay test
- conditions
- strain
Senders:XL10GOLD
Receiver:JW1908
- temparature
30°C and Room Temparature
Team:Output
colony PCR
ptet+LuxR(×16)
ptet+LasR(×7)
ptet+RhlR(×5)
ptet+cinR(×5)
