Team:Chiba/Calendar-Home/20 August 2008

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(Difference between revisions)

Revision as of 06:55, 24 October 2008

>Home | Notebook

19 August 2008 <|> 21 August 2008

Laboratory work

Team:Input

PCR

[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2007)①
[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2008)②
[http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2007)③
[http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2008)④
[http://partsregistry.org/Part:BBa_E0612 BBa_E0612](2008)⑤

>

Sample No.	①	②	③	④	⑤
DNA tamplate	1	1	1	1	1
FW primer	5	5	5	5	5
VR primer	5	5	5	5	5
dNTPmix	10	10	10	10	10
thermo pol buffer	10	10	10	10	10
DNA pol(VENT)	1	1	1	1	1
dH₂O	68	68	68	68	68
TOTAL	100	100	100	100	100

-->Gel Check

>

Sample No.	①	②	③	④	⑤
Sample DNA	1	1	1	1	1
loading dye	1	1	1	1	1
dH₂O	4	4	4	4	4
Total	6	6	6	6	6

-->どれもバンド出ず。

Transformation

[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)
[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2006)
[http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2007)
[http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2006)
[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2007)
[http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2007)

-->BBa_J22141のみコロニーがほとんど生えていなかった。(１個のみ)

Team:Communication

(18/8)--->Mini prep

BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070]
BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076]
BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]
BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]

--->Gel Check

Sample DNA	3
Loading Dye	2
dH2O	7
TOTAL	12μL

From left;

BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None
BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None
BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> None

Sample DNA	10
Loading Dye	4
dH2O	10
TOTAL	24μL

From left;

BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None
BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> OK
BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None
BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070] -> OK
BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070] -> OK

Sample DNA	3
Loading Dye	2
dH2O	7
TOTAL	12μL

From left;

BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK
BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK
BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK
BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK

--->Digestion test

BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]

Sample No	1	2	3	4	5	6	7	8
BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]	3	-	-	-	3	-	-	-
BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]	-	3	-	-	-	3	-	-
BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]	-	-	3	-	-	-	3	-
BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]	-	-	-	3	-	-	-	3
EcoRⅠ	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.1
PstⅠ	-	-	-	-	0.1	0.1	0.1	0.1
Buffer EcoRⅠ	1	1	1	1	1	1	1	1
BSA	1	1	1	1	1	1	1	1
dH2O	4.9	4.9	4.9	4.9	4.8	4.8	4.8	4.8
TOTAL	10	10	10	10	10	10	10	10

--->Gel Check

Sample DNA	3
Loading Dye	2
dH2O	7
TOTAL	12μL

From left;

(Single Digestion 1~4)

BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK
BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK
BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK
BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK

(Double Digestion 1~4)

BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK
BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK
BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK
BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK

Team:Output

Transformation

[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](Luciferase Renilla)-->colony
[http://partsregistry.org/Part:BBa_I712019 BBa_I712019](Luciferase Fire fly)-->no colony
[http://partsregistry.org/Part:BBa_J33202 BBa_J33202](LacZ)-->colony
[http://partsregistry.org/Part:BBa_E2030 BBa_E2030](Venus YFP)-->not so good
YFP(P-GEX)-->colony
[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]-->colony
[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]-->colony

Luciferase(J52008:2007,2008 I712019:2008)(Amp)
Lac Z(J33202:2007,2008)(Amp)
Venus(E2030:2007,2008)(Kam)
Craig Venus(Amp?Kan?)
T9002(Amp)
F2620(Amp)

luciferase,LacZ,Venus
competent cell 50μl
DNA 1μl
soc 200μl
plating 250μl

Craig Venus,T9002,F2620
competent cell 30μl
DNA 1μl
soc 120μl
plating 20μl

result

成功→Luciferase(J52008:2007),Lac Z(J33202:2007),Craig Venus,T9002,F2620

失敗→Luciferase(J52008:2008 J52008:2008),Lac Z(J33202:2008),Venus(J32007:2007,2008)

@@ Line 1: / Line 1: @@
-'''OUTPUT TEAM'''
+>[[Team:Chiba|Home]] | [[Team:Chiba/Notebook|Notebook]]
-<br>Transformation
+[[Team:Chiba/Calendar-Home/19 August 2008|19 August 2008 <]]|[[Team:Chiba/Calendar-Home/21 August 2008|> 21 August 2008]]
+==Laboratory work==
+===Team:Input===
+'''[[Team:Chiba/protocol/PCR|PCR]]'''
+*[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2007)①
+*[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2008)②
+*[http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2007)③
+*[http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2008)④
+*[http://partsregistry.org/Part:BBa_E0612 BBa_E0612](2008)⑤
+<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<td width="257">Sample No.</td>
+<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
+</tr>
+<tr>
+<td>DNA tamplate</td>
+<td>1</td><td>1</td><td>1</td><td>1</td>><td>1</td>
+</tr>
+<tr>
+<td>FW primer</td>
+<td>5</td><td>5</td><td>5</td><td>5</td><td>5</td>
+</tr>
+<tr>
+<td>VR primer</td>
+<td>5</td><td>5</td><td>5</td><td>5</td><td>5</td>
+</tr>
+<tr>
+<td>dNTPmix</td>
+<td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
+</tr>
+<tr>
+<td>thermo pol buffer</td>
+<td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
+</tr>
+<tr>
+<td>DNA pol(VENT)</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>68</td><td>68</td><td>68</td><td>68</td><td>68</td>
+</tr>
+<tr>
+<td>TOTAL</td>
+<td>100</td><td>100</td><td>100</td><td>100</td><td>100</td>
+</tr>
+</table>
+-->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
+<table width="200" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<td width="257">Sample No.</td>
+<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
+</tr>
+<tr>
+<td>Sample DNA</td>
+<td>1</td><td>1</td><td>1</td><td>1</td>><td>1</td>
+</tr>
+<tr>
+<td>loading dye</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>4</td><td>4</td><td>4</td><td>4</td><td>4</td>
+</tr>
+<tr>
+<td>Total</td>
+<td>6</td><td>6</td><td>6</td><td>6</td><td>6</td>
+</tr>
+</table>
+-->どれもバンド出ず。
+'''[[Team:Chiba/protocol/transformation|Transformation]]'''
+*[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)
+*[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2006)
+*[http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2007)
+*[http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2006)
+*[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2007)
+*[http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2007)
+-->BBa_J22141のみコロニーがほとんど生えていなかった。(１個のみ)
+===Team:Communication===
+:(18/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
+::*BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
+::*BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070]
+::*BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076]
+::*BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
+::*BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]
+::*BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
+::*BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
+::*BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
+::*BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]
+:--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
+{|align="justify"
+|[[Image:Chiba-0820-3.JPG]]
+|
+:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<td width="257">Sample DNA</td>
+<td>3</td>
+</tr>
+<tr>
+<td>Loading Dye</td>
+<td>2</td>
+</tr>
+<tr>
+<td>dH2O</td>
+<td>7</td>
+</tr>
+<tr>
+<td>TOTAL</td>
+<td>12μL</td>
+</tr>
+</table>
+:From left;
+::*BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None
+::*BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None
+::*BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> None
+|}
+{|align="justify"
+|[[Image:Chiba-0820-4.JPG]]
+|
+:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<td width="257">Sample DNA</td>
+<td>10</td>
+</tr>
+<tr>
+<td>Loading Dye</td>
+<td>4</td>
+</tr>
+<tr>
+<td>dH2O</td>
+<td>10</td>
+</tr>
+<tr>
+<td>TOTAL</td>
+<td>24μL</td>
+</tr>
+</table>
+:From left;
+::*BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None
+::*BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> OK
+::*BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None
+::*BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070] -> OK
+::*BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070] -> OK
+|}
+{|align="justify"
+|[[Image:Chiba-0820.JPG]]
+|
+:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<td width="257">Sample DNA</td>
+<td>3</td>
+</tr>
+<tr>
+<td>Loading Dye</td>
+<td>2</td>
+</tr>
+<tr>
+<td>dH2O</td>
+<td>7</td>
+</tr>
+<tr>
+<td>TOTAL</td>
+<td>12μL</td>
+</tr>
+</table>
+:From left;<br>
+::*BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK<br>
+::*BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK<br>
+::*BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK<br>
+::*BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK<br>
+|}
+--->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
+::*BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
+::*BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
+::*BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
+::*BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]
+<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<td width="257">Sample No</td>
+<td>1</td><td>2</td><td>3</td><td>4</td><td>5</td><td>6</td><td>7</td><td>8</td>
+</tr>
+<tr>
+<td>BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]</td>
+<td>3</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td>
+</tr>
+<tr>
+<td>BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]</td>
+<td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td>
+</tr>
+<tr>
+<td>BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]</td>
+<td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td>
+</tr>
+<tr>
+<td>BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]</td>
+<td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>3</td>
+</tr>
+<tr>
+<td>EcoRⅠ</td>
+<td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
+</tr>
+<tr>
+<td>PstⅠ</td>
+<td>-</td><td>-</td><td>-</td><td>-</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
+</tr>
+<tr>
+<td>Buffer EcoRⅠ</td>
+<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
+</tr>
+<tr>
+<td>BSA</td>
+<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
+</tr>
+<tr>
+<td>dH2O</td>
+<td>4.9</td><td>4.9</td><td>4.9</td><td>4.9</td><td>4.8</td><td>4.8</td><td>4.8</td><td>4.8</td>
+</tr>
+<tr>
+<td>TOTAL</td>
+<td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
+</tr>
+</table>
+:--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
+{|align="justify"
+|[[Image:Chiba-0820-2.JPG]]
+|
+:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<td width="257">Sample DNA</td>
+<td>3</td>
+</tr>
+<tr>
+<td>Loading Dye</td>
+<td>2</td>
+</tr>
+<tr>
+<td>dH2O</td>
+<td>7</td>
+</tr>
+<tr>
+<td>TOTAL</td>
+<td>12μL</td>
+</tr>
+</table>
+:From left;
+:(Single Digestion 1~4)
+::*BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK
+::*BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK
+::*BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK
+::*BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK
+:(Double Digestion 1~4)
+::*BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK
+::*BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK
+::*BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK
+::*BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK
+|}
+===Team:Output===
+[[Team:Chiba/protocol/transformation|Transformation]]
+*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](Luciferase Renilla)-->colony
+*[http://partsregistry.org/Part:BBa_I712019 BBa_I712019](Luciferase Fire fly)-->no colony
+*[http://partsregistry.org/Part:BBa_J33202 BBa_J33202](LacZ)-->colony
+*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030](Venus YFP)-->not so good
+*YFP(P-GEX)-->colony
+*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]-->colony
+*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]-->colony
 <br>Luciferase(J52008:2007,2008 I712019:2008)(Amp)
 <br>Lac Z(J33202:2007,2008)(Amp)
@@ Line 7: / Line 310: @@
 <br>T9002(Amp)
 <br>F2620(Amp)
 <br>luciferase,LacZ,Venus

Team:Chiba/Calendar-Home/20 August 2008

From 2008.igem.org

Revision as of 06:55, 24 October 2008

Contents

Laboratory work

Team:Input

Team:Communication

Team:Output