Team:Chiba/Calendar-Home/25 September 2008
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===Team:Output=== | ===Team:Output=== | ||
- | <br>colony PCR | + | <br>colony [[Team:Chiba/protocol/PCR|PCR]] |
- | <br>Vector:BBa_R0062 Insert:BBa_E0240 | + | <br>Vector:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] Insert:[http://partsregistry.org/Part:BBa_E0240 BBa_E0240] |
- | <br>Vector:BBa_R0062 | + | <br>Vector:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] |
<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>each</td> | <td>each</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>culture</td> | + | <td>culture(μL)</td> |
<td>1</td> | <td>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Fwd primer</td> | + | <td>Fwd primer(μL)</td> |
<td>1.5</td> | <td>1.5</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Rev primer</td> | + | <td>Rev primer(μL)</td> |
<td>1.5</td> | <td>1.5</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Taq(DNA pol)</td> | + | <td>Taq(DNA pol)(μL)</td> |
<td>0.2</td> | <td>0.2</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>dNTPmix</td> | + | <td>dNTPmix(μL)</td> |
<td>3</td> | <td>3</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>thermo pol Buffer</td> | + | <td>thermo pol Buffer(μL)</td> |
<td>3</td> | <td>3</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O(μL)</td> |
<td>19.8</td> | <td>19.8</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>TOTAL</td> | + | <td>TOTAL(μL)</td> |
<td>30</td> | <td>30</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | <br>-> | + | <br>->95°C 5 min ->(95°C 1 min 50°C 30sec 72°C 1 min)×25 ->72°C 10 min |
- | <br>Digestion | + | <br>[[Team:Chiba/protocol/digestion|Digestion test]] |
- | <br>BBa_I6206 | + | <br>[http://partsregistry.org/Part:BBa_I6206 BBa_I6206] |
- | <br>BBa_I6101 | + | <br>[http://partsregistry.org/Part:BBa_I6101 BBa_I6101] |
- | <br>BBa_I6042 | + | <br>[http://partsregistry.org/Part:BBa_I6042 BBa_I6042] |
- | <br>BBa_I0462 | + | <br>[http://partsregistry.org/Part:BBa_I0462 BBa_I0462] |
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
Line 62: | Line 62: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>DNA</td> | + | <td>DNA(μL)</td> |
<td>2</td> | <td>2</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>EcoRI(μL)</td> |
<td>0.2</td> | <td>0.2</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>EcoRI Buffer(μL)</td> |
<td>1</td> | <td>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O(μL)</td> |
<td>7</td> | <td>7</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>TOTAL</td> | + | <td>TOTAL(μL)</td> |
<td>30</td> | <td>30</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | <br>-> | + | <br>->30 min 37°C |
- | <br> | + | <br>[[Team:Chiba/protocol/gelcheck|gelcheck]] |
- | <br>BBa_I6206(digestion product) | + | <br>[http://partsregistry.org/Part:BBa_I6206 BBa_I6206](digestion product) |
- | <br>BBa_I6101(digestion product) | + | <br>[http://partsregistry.org/Part:BBa_I6101 BBa_I6101](digestion product) |
- | <br>BBa_I6042(digestion product) | + | <br>[http://partsregistry.org/Part:BBa_I6042 BBa_I6042](digestion product) |
- | <br>BBa_I0462(digestion product) | + | <br>[http://partsregistry.org/Part:BBa_I0462 BBa_I0462](digestion product) |
- | <br>BBa_I0642(natural) | + | <br>[http://partsregistry.org/Part:BBa_I0642 BBa_I0642](natural) |
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
Line 96: | Line 96: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>DNA</td> | + | <td>DNA(μL)</td> |
<td>10</td><td>1</td> | <td>10</td><td>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Loading Dye</td> | + | <td>Loading Dye(μL)</td> |
<td>2</td><td>1</td> | <td>2</td><td>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O(μL)</td> |
<td>-</td><td>4</td> | <td>-</td><td>4</td> | ||
<tr> | <tr> | ||
- | <td>TOTAL</td> | + | <td>TOTAL(μL)</td> |
<td>12</td><td>6</td> | <td>12</td><td>6</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | <br> | + | <br>[[Team:Chiba/protocol/digestion|digestion]] |
- | <br>BBa_I0462 | + | <br>[http://partsregistry.org/Part:BBa_I0462 BBa_I0462] |
<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">DNA</td> | + | <td width="257">DNA(μL)</td> |
<td>25</td> | <td>25</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>XbaI(μL)</td> |
<td>1</td> | <td>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>PstI</td> |
<td>1</td> | <td>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Buffer 3</td> | + | <td>Buffer 3(μL)</td> |
<td>4</td> | <td>4</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>BSA</td> | + | <td>BSA(μL)</td> |
<td>4</td> | <td>4</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O(μL)</td> |
<td>5</td> | <td>5</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>TOTAL</td> | + | <td>TOTAL(μL)</td> |
<td>40</td> | <td>40</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | <br>-> | + | <br>->3 hours 37°C |
- | <br>->Zymo -> | + | <br>->Zymo ->10μL |
- | <br>Ligation | + | <br>[[Team:Chiba/protocol/ligation/ligation|Ligation]] |
- | <br>Vector:BBa_I7100 | + | <br>Vector:[http://partsregistry.org/Part:BBa_I7100 BBa_I7100] |
- | <br>Insert:BBa_R0062 | + | <br>Insert:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] |
- | <br>Insert:BBa_I0462 | + | <br>Insert:[http://partsregistry.org/Part:BBa_I0462 BBa_I0462] |
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
Line 158: | Line 158: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>[http://partsregistry.org/Part:BBa_R0062 BBa_R0062]</td> |
<td>2</td><td>-</td> | <td>2</td><td>-</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>[http://partsregistry.org/Part:BBa_I0462 BBa_I0462]</td> |
<td>3.3</td><td>-</td> | <td>3.3</td><td>-</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Ligase</td> | + | <td>Ligase(μL)</td> |
<td>1</td><td>1</td> | <td>1</td><td>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>Ligase Buffer</td> | + | <td>Ligase Buffer(μL)</td> |
<td>2</td><td>2</td> | <td>2</td><td>2</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O(μL)</td> |
<td>0.4</td><td>5</td> | <td>0.4</td><td>5</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>TOTAL</td> | + | <td>TOTAL(μL)</td> |
<td>10</td><td>10</td> | <td>10</td><td>10</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | <br>-> | + | <br>->3 hours 37°C |
- | <br>->Transformation | + | <br>->[[Team:Chiba/protocol/transformation|Transformation]] |
Latest revision as of 23:29, 29 October 2008
24 September 2008 <|> 26 September 2008
Laboratory work
Team:Communication
Team:Output
colony PCR
Vector:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] Insert:[http://partsregistry.org/Part:BBa_E0240 BBa_E0240]
Vector:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062]
Sample No. | each |
culture(μL) | 1 |
Fwd primer(μL) | 1.5 |
Rev primer(μL) | 1.5 |
Taq(DNA pol)(μL) | 0.2 |
dNTPmix(μL) | 3 |
thermo pol Buffer(μL) | 3 |
dH2O(μL) | 19.8 |
TOTAL(μL) | 30 |
->95°C 5 min ->(95°C 1 min 50°C 30sec 72°C 1 min)×25 ->72°C 10 min
Digestion test
[http://partsregistry.org/Part:BBa_I6206 BBa_I6206]
[http://partsregistry.org/Part:BBa_I6101 BBa_I6101]
[http://partsregistry.org/Part:BBa_I6042 BBa_I6042]
[http://partsregistry.org/Part:BBa_I0462 BBa_I0462]
Sample No | each |
DNA(μL) | 2 |
EcoRI(μL) | 0.2 |
EcoRI Buffer(μL) | 1 |
dH2O(μL) | 7 |
TOTAL(μL) | 30 |
->30 min 37°C
gelcheck
[http://partsregistry.org/Part:BBa_I6206 BBa_I6206](digestion product)
[http://partsregistry.org/Part:BBa_I6101 BBa_I6101](digestion product)
[http://partsregistry.org/Part:BBa_I6042 BBa_I6042](digestion product)
[http://partsregistry.org/Part:BBa_I0462 BBa_I0462](digestion product)
[http://partsregistry.org/Part:BBa_I0642 BBa_I0642](natural)
Sample No | each(digestion) | I0462(natural) |
DNA(μL) | 10 | 1 |
Loading Dye(μL) | 2 | 1 |
dH2O(μL) | - | 4 |
TOTAL(μL) | 12 | 6 |
digestion
[http://partsregistry.org/Part:BBa_I0462 BBa_I0462]
DNA(μL) | 25 |
XbaI(μL) | 1 |
PstI | 1 |
Buffer 3(μL) | 4 |
BSA(μL) | 4 |
dH2O(μL) | 5 |
TOTAL(μL) | 40 |
->3 hours 37°C
->Zymo ->10μL
Ligation
Vector:[http://partsregistry.org/Part:BBa_I7100 BBa_I7100]
Insert:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062]
Insert:[http://partsregistry.org/Part:BBa_I0462 BBa_I0462]
I7100 | 1.3 | 1.3 |
[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] | 2 | - |
[http://partsregistry.org/Part:BBa_I0462 BBa_I0462] | 3.3 | - |
Ligase(μL) | 1 | 1 |
Ligase Buffer(μL) | 2 | 2 |
dH2O(μL) | 0.4 | 5 |
TOTAL(μL) | 10 | 10 |
->3 hours 37°C
->Transformation