Team:Chiba/Calendar-Home/25 August 2008

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(Difference between revisions)

Latest revision as of 06:15, 30 October 2008

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24 August 2008 <|> 26 August 2008

Laboratory work

Team:Input

The following glycerol Stock was made.

BBa_J22136(insert check OK)

Picked the colony and incubate with 2mL of LB-Ampicillin Medium for 12 hours at 37 degrees. Took 180μl from culture,and mixed with 880μlLB-Amp Medium.(OD=0.485)

Liquid culture(OD=0.495)	0.67μl
60%Glycerol	0.33μl
20%Glycerol Stock	1.00ml

Team:Communication

(24/8)--->Digestion

Double Digestion:BBa_J04500(2007)
Double Digesiton:BBa_R0010(2007)
Single Digestion:BBa_R0010(2007)
BBa_R0010(2007)

Sample No.	1	2	3	4
Sample DNA(μL)	5	5	1	1
EcoRI(μL)	0.5	0.5	-	-
SpeI(μL)	0.5	0.5	-	-
Buffer 1	1	1	-	-
Buffer 3(μL)	-	-	1	-
BSA(μL)	1	1	-	-
dH₂O(μL)	2	2	7.5	-
TOTAL(μL)	10	10	10	1

--->Gel Check

Sample No.	1~3	4
Sample DNA(μL)	10	1
Loading Dye(μL)	2	1
dH₂O(μL)	-	4
TOTAL(μL)	12	6

From right;

Double Digestion:BBa_J04500(2007) -> OK
Double Digesiton:BBa_R0010(2007) -> ?(low)
Single Digestion:BBa_R0010(2007) -> ?(low)
BBa_R0010(2007) -> OK(low)

--->Digestion (Double Digestion)

Sample No.	1	2	3
Sample DNA(μL)	33ng/μl×60μl	33ng/μl×60μl	100ng/μl×20μl
PstI(μL)	2	2	2
XbaI(μL)	2	2	-
SpeI(μL)	-	-	2
Buffer 2(μL)	-	-	5
Buffer 3(μL)	10	10	-
BSA(μL)	10	10	5
dH₂O(μL)	16	16	16
TOTAL(μL)	100	100	52

Team:Output

PCR

No.	1	2	3	4	5
DNA tamplate(μL)	1	1	1	1	1
FW primer(μL)	5(Venus_YFP_fwd)	5(LacZ_fwd)	5(LacZ_fwd)	5(rLUC_fwd)	5(fLuc_fwd)
VR primer(μL)	5(VR)	5(LacZ_rev)	5(LacZ_rev)	5(VR)	5(VR)
dNTPmix(μL)	10	10	10	10	10
Thermo pol buffer(μL)	10	10	10	10	10
DNA pol(VENT)(μL)	1	1	1	1	1
dH₂O(μL)	68	68	68	68	68
TOTAL(μL)	100	100	100	100	100

-->Gel Check

No.	1	2	3	4	5
DNA tamplate(μL)	1	1	1	1	1
loading Dye(μL)	1	1	1	1	1
dH₂O(μL)	4	4	4	4	4
TOTAL(μL)	6	6	6	6	6

Mini prep

Transformation

BBa_E2030
PUC19
pGFPUV
pGEX Venus YFP
Membren Venus YFP

PCR

BBa_J63001(1)

No.	1
DNA tamplate(μL)	1
FW primer(Venus)(μL)	5
VR primer(μL)	5
dNTPmix(μL)	10
Thermo pol buffer(μL)	10
DNA pol(VENT)(μL)	1
dH₂O(μL)	68
TOTAL(μL)	100

@@ Line 168: / Line 168: @@
 </tr>
 <tr>
-<td>DNA tamplate</td>
+<td>DNA tamplate(&mu;L)</td>
 <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
 </tr>
 <tr>
-<td>FW primer</td>
+<td>FW primer(&mu;L)</td>
 <td>5(Venus_YFP_fwd)</td><td>5(LacZ_fwd)</td><td>5(LacZ_fwd)</td><td>5(rLUC_fwd)</td><td>5(fLuc_fwd)</td>
 </tr>
 <tr>
-<td>VR primer</td>
+<td>VR primer(&mu;L)</td>
 <td>5(VR)</td><td>5(LacZ_rev)</td><td>5(LacZ_rev)</td><td>5(VR)</td><td>5(VR)</td>
 </tr>
 <tr>
-<td>dNTPmix</td>
+<td>dNTPmix(&mu;L)</td>
 <td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
 </tr>
 <tr>
-<td>Thermo pol buffer</td>
+<td>Thermo pol buffer(&mu;L)</td>
 <td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
 </tr>
 <tr>
-<td>DNA pol(VENT)</td>
+<td>DNA pol(VENT)(&mu;L)</td>
 <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
 </tr>
 <tr>
-<td>dH<sub>2</sub>O</td>
+<td>dH<sub>2</sub>O(&mu;L)</td>
 <td>68</td><td>68</td><td>68</td><td>68</td><td>68</td>
 </tr>
 <tr>
-<td>TOTAL</td>
+<td>TOTAL(&mu;L)</td>
-<td>100μl</td><td>100μl</td><td>100μl</td><td>100μl</td><td>100μl</td>
+<td>100</td><td>100</td><td>100</td><td>100</td><td>100</td>
 </tr>
 </table>
@@ Line 210: / Line 210: @@
 </tr>
 <tr>
-<td>DNA tamplate</td>
+<td>DNA tamplate(&mu;L)</td>
 <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
 </tr>
 <tr>
-<td>loading Dye</td>
+<td>loading Dye(&mu;L)</td>
 <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
 </tr>
 <tr>
-<td>dH<sub>2</sub>O</td>
+<td>dH<sub>2</sub>O(&mu;L)</td>
 <td>4</td><td>4</td><td>4</td><td>4</td><td>4</td>
 </tr>
 <tr>
-<td>TOTAL</td>
+<td>TOTAL(&mu;L)</td>
-<td>6μl</td><td>6μl</td><td>6μl</td><td>6μl</td><td>6μl</td>
+<td>6</td><td>6</td><td>6</td><td>6</td><td>6</td>
 </tr>
 </table>
@@ Line 246: / Line 246: @@
 </tr>
 <tr>
-<td>DNA tamplate</td>
+<td>DNA tamplate(&mu;L)</td>
 <td>1</td>
 </tr>
 <tr>
-<td>FW primer(Venus)</td>
+<td>FW primer(Venus)(&mu;L)</td>
 <td>5</td>
 </tr>
 <tr>
-<td>VR primer</td>
+<td>VR primer(&mu;L)</td>
 <td>5</td>
 </tr>
 <tr>
-<td>dNTPmix</td>
+<td>dNTPmix(&mu;L)</td>
 <td>10</td>
 </tr>
 <tr>
-<td>Thermo pol buffer</td>
+<td>Thermo pol buffer(&mu;L)</td>
 <td>10</td>
 </tr>
 <tr>
-<td>DNA pol(VENT)</td>
+<td>DNA pol(VENT)(&mu;L)</td>
 <td>1</td>
 </tr>
 <tr>
-<td>dH<sub>2</sub>O</td>
+<td>dH<sub>2</sub>O(&mu;L)</td>
 <td>68</td>
 </tr>
 <tr>
-<td>TOTAL</td>
+<td>TOTAL(&mu;L)</td>
-<td>100μl</td>
+<td>100</td>
 </tr>
 </table>