Team:Chiba/Calendar-Home/25 September 2008

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Revision as of 20:10, 29 October 2008

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24 September 2008 <|> 26 September 2008

Laboratory work

Team:Communication

Team:Output

colony PCR
Vector:BBa_R0062 Insert:BBa_E0240
Vector: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
BBa_I6206
BBa_I6101
BBa_I6042
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
Gel check
BBa_I6206(digestion product)
BBa_I6101(digestion product)
BBa_I6042(digestion product)
BBa_I0462(digestion product)
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
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:BBa_I7100
Insert:BBa_R0062
Insert:BBa_I0462

I7100	1.3	1.3
R0062	2	-
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

@@ Line 12: / Line 12: @@
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No</td>
+<td width="257">Sample No.</td>
 <td>each</td>
 </tr>
 <tr>
-<td>culture</td>
+<td>culture(&mu;L)</td>
 <td>1</td>
 </tr>
 <tr>
-<td>Fwd primer</td>
+<td>Fwd primer(&mu;L)</td>
 <td>1.5</td>
 </tr>
 <tr>
-<td>Rev primer</td>
+<td>Rev primer(&mu;L)</td>
 <td>1.5</td>
 </tr>
 <tr>
-<td>Taq(DNA pol)</td>
+<td>Taq(DNA pol)(&mu;L)</td>
 <td>0.2</td>
 </tr>
 <tr>
-<td>dNTPmix</td>
+<td>dNTPmix(&mu;L)</td>
 <td>3</td>
 </tr>
 <tr>
-<td>thermo pol Buffer</td>
+<td>thermo pol Buffer(&mu;L)</td>
 <td>3</td>
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O(&mu;L)</td>
 <td>19.8</td>
 </tr>
 <tr>
-<td>TOTAL</td>
+<td>TOTAL(&mu;L)</td>
 <td>30</td>
 </tr>
 </table>
-<br>->95℃ 5min ->(95℃ 1min 50℃ 30sec　72℃ 1min)x25 ->72℃ 10min
+<br>->95&deg;C 5 min ->(95&deg;C 1 min 50&deg;C 30sec　72&deg;C 1 min)&times;25 ->72&deg;C 10 min
 <br>Digestion Test
@@ Line 62: / Line 62: @@
 </tr>
 <tr>
-<td>DNA</td>
+<td>DNA(&mu;L)</td>
 <td>2</td>
 </tr>
 <tr>
-<td>EcoRⅠ</td>
+<td>EcoRI(&mu;L)</td>
 <td>0.2</td>
 </tr>
 <tr>
-<td>EcoRⅠ Buffer</td>
+<td>EcoRI Buffer(&mu;L)</td>
 <td>1</td>
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O(&mu;L)</td>
 <td>7</td>
 </tr>
 <tr>
-<td>TOTAL</td>
+<td>TOTAL(&mu;L)</td>
 <td>30</td>
 </tr>
 </table>
-<br>->30min 37℃
+<br>->30 min 37&deg;C
-<br>Gel cheack
+<br>Gel check
 <br>BBa_I6206(digestion product)
 <br>BBa_I6101(digestion product)
@@ Line 96: / Line 96: @@
 </tr>
 <tr>
-<td>DNA</td>
+<td>DNA(&mu;L)</td>
 <td>10</td><td>1</td>
 </tr>
 <tr>
-<td>Loading Dye</td>
+<td>Loading Dye(&mu;L)</td>
 <td>2</td><td>1</td>
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O(&mu;L)</td>
 <td>-</td><td>4</td>
 <tr>
-<td>TOTAL</td>
+<td>TOTAL(&mu;L)</td>
 <td>12</td><td>6</td>
 </tr>
@@ Line 116: / Line 116: @@
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">DNA</td>
+<td width="257">DNA(&mu;L)</td>
 <td>25</td>
 </tr>
 <tr>
-<td>XbaⅠ</td>
+<td>XbaI(&mu;L)</td>
 <td>1</td>
 </tr>
 <tr>
-<td>PstⅠ</td>
+<td>PstI</td>
 <td>1</td>
 </tr>
 <tr>
-<td>Buffer 3</td>
+<td>Buffer 3(&mu;L)</td>
 <td>4</td>
 </tr>
 <tr>
-<td>BSA</td>
+<td>BSA(&mu;L)</td>
 <td>4</td>
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O(&mu;L)</td>
 <td>5</td>
 </tr>
 <tr>
-<td>TOTAL</td>
+<td>TOTAL(&mu;L)</td>
 <td>40</td>
 </tr>
 </table>
-<br>->3hour 37℃
+<br>->3 hours 37&deg;C
-<br>->Zymo ->10μl
+<br>->Zymo ->10&mu;L
 <br>Ligation
@@ Line 166: / Line 166: @@
 </tr>
 <tr>
-<td>Ligase</td>
+<td>Ligase(&mu;L)</td>
 <td>1</td><td>1</td>
 </tr>
 <tr>
-<td>Ligase Buffer</td>
+<td>Ligase Buffer(&mu;L)</td>
 <td>2</td><td>2</td>
 </tr>
 <tr>
-<td>dH2O
+<td>dH<sub>2</sub>O(&mu;L)<td>
 <td>0.4</td><td>5</td>
 </tr>
 <tr>
-<td>TOTAL</td>
+<td>TOTAL(&mu;L)</td>
 <td>10</td><td>10</td>
 </tr>
 </table>
-<br>->3hour 37℃
+<br>->3 hours 37&deg;C
 <br>->Transformation