Team:Chiba/Calendar-Home/25 August 2008

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(Difference between revisions)

Latest revision as of 06:15, 30 October 2008

>Home | Notebook

24 August 2008 <|> 26 August 2008

Laboratory work

Team:Input

The following glycerol Stock was made.

[http://partsregistry.org/Part:BBa_J22136 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:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007)
Double Digesiton:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
Single Digestion:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
[http://partsregistry.org/Part:BBa_R0010 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:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007) -> OK
Double Digesiton:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007) -> ?(low)
Single Digestion:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007) -> ?(low)
[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007) -> OK(low)

--->Digestion (Double Digestion)

[http://partsregistry.org/Part:BBa_C0178 BBa_C0178]
[http://partsregistry.org/Part:BBa_C0170 BBa_C0170]
[http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007)

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

[http://partsregistry.org/Part:BBa_E2030 BBa_E2030](1)
[http://partsregistry.org/Part:BBa_J33202 BBa_J33202](2)
BL21(3)
[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](4)
[http://partsregistry.org/Part:BBa_I712019 BBa_I712019](5)

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

[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]

Transformation

[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
PUC19
pGFPUV
pGEX Venus YFP
Membren Venus YFP

PCR

[http://partsregistry.org/Part:BBa_J63001 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 7: / Line 7: @@
 The following glycerol Stock was made.
-[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](insert check済み)
+[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](insert check OK)
-プレートに生えたコロニーをpickして、試験管で2ml培養(LB-Amp,37℃,12h)
+<BR>Picked the colony and incubate with 2mL of LB-Ampicillin Medium for 12 hours at 37 degrees.
+Took 180&mu;l from culture,and mixed with 880&mu;lLB-Amp Medium.(OD=0.485)
-培養した試験管から180μl,LB-Ampを880μlとって、OD=0.495にした。
 <table width="210" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
@@ Line 39: / Line 38: @@
 </tr>
 <tr>
-<td>Sample DNA</td>
+<td>Sample DNA(&mu;L)</td>
 <td>5</td><td>5</td><td>1</td><td>1</td>
 </tr>
 <tr>
-<td>EcoRⅠ</td>
+<td>EcoRI(&mu;L)</td>
 <td>0.5</td><td>0.5</td><td>-</td><td>-</td>
 </tr>
 <tr>
-<td>SpeⅠ</td>
+<td>SpeI(&mu;L)</td>
 <td>0.5</td><td>0.5</td><td>-</td><td>-</td>
 </tr>
@@ Line 55: / Line 54: @@
 </tr>
 <tr>
-<td>Buffer 3</td>
+<td>Buffer 3(&mu;L)</td>
 <td>-</td><td>-</td><td>1</td><td>-</td>
 </tr>
 <tr>
-<td>BSA</td>
+<td>BSA(&mu;L)</td>
 <td>1</td><td>1</td><td>-</td><td>-</td>
 </tr>
 <tr>
-<td>dH<sub>2</sub>O</td>
+<td>dH<sub>2</sub>O(&mu;L)</td>
 <td>2</td><td>2</td><td>7.5</td><td>-</td>
 </tr>
 <tr>
-<td>TOTAL</td>
+<td>TOTAL(&mu;L)</td>
 <td>10</td><td>10</td><td>10</td><td>1</td>
 </tr>
@@ Line 82: / Line 81: @@
 </tr>
 <tr>
-<td>Sample DNA</td>
+<td>Sample 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>dH<sub>2</sub>O</td>
+<td>dH<sub>2</sub>O(&mu;L)</td>
 <td>-</td><td>4</td>
 </tr>
 <tr>
-<td>TOTAL</td>
+<td>TOTAL(&mu;L)</td>
 <td>12</td><td>6</td>
 </tr>
@@ Line 118: / Line 117: @@
 </tr>
 <tr>
-<td>Sample DNA</td>
+<td>Sample DNA(&mu;L)</td>
 <td>33ng/μl×60μl</td><td>33ng/μl×60μl</td><td>100ng/μl×20μl</td>
 </tr>
 <tr>
-<td>PstⅠ</td>
+<td>PstI(&mu;L)</td>
 <td>2</td><td>2</td><td>2</td>
 </tr>
 <tr>
-<td>XbaⅠ</td>
+<td>XbaI(&mu;L)</td>
 <td>2</td><td>2</td><td>-</td>
 </tr>
 <tr>
-<td>SpeⅠ</td>
+<td>SpeI(&mu;L)</td>
 <td>-</td><td>-</td><td>2</td>
 </tr>
 <tr>
-<td>Buffer 2</td>
+<td>Buffer 2(&mu;L)</td>
 <td>-</td><td>-</td><td>5</td>
 </tr>
 <tr>
-<td>Buffer 3</td>
+<td>Buffer 3(&mu;L)</td>
 <td>10</td><td>10</td><td>-</td>
 </tr>
 <tr>
-<td>BSA</td>
+<td>BSA(&mu;L)</td>
 <td>10</td><td>10</td><td>5</td>
 </tr>
 <tr>
-<td>dH<sub>2</sub>O</td>
+<td>dH<sub>2</sub>O(&mu;L)</td>
 <td>16</td><td>16</td><td>16</td>
 </tr>
 <tr>
-<td>TOTAL</td>
+<td>TOTAL(&mu;L)</td>
 <td>100</td><td>100</td><td>52</td>
 </tr>
 </table>
 ===Team:Output===
 [[Team:Chiba/protocol/PCR|PCR]]
-*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]①
+*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030](1)
-*[http://partsregistry.org/Part:BBa_J33202 BBa_J33202]②
+*[http://partsregistry.org/Part:BBa_J33202 BBa_J33202](2)
-*BL21③
+*BL21(3)
-*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]④
+*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](4)
-*[http://partsregistry.org/Part:BBa_I712019 BBa_I712019]⑤
+*[http://partsregistry.org/Part:BBa_I712019 BBa_I712019](5)
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No.</td>
+<td width="257">No.</td>
-<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
+<td>1</td><td>2</td><td>3</td><td>4</td><td>5</td>
 </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 209: / Line 206: @@
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No.</td>
+<td width="257">No.</td>
-<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
+<td>1</td><td>2</td><td>3</td><td>4</td><td>5</td>
 </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>
 [[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]]
-*Venus
+*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
 *[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
 [[Team:Chiba/protocol/transformation|Transformation]]
-*Venus
+*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
 *PUC19
 *pGFPUV
@@ Line 242: / Line 239: @@
 *Membren Venus YFP
 [[Team:Chiba/protocol/PCR|PCR]]
-*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001]①
+*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001](1)
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No.</td>
+<td width="257">No.</td>
-<td>①</td>
+<td>1</td>
 </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>