Team:Chiba/jk/γ/week 3

From 2008.igem.org

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<td>1</td>
<td>1</td>
</tr>
</tr>
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<td>Fwd primer</td>
<td>Fwd primer</td>
<td>1.5</td>
<td>1.5</td>
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== 3,September,2008 ==
== 3,September,2008 ==
-
[[Team:Chiba/protocol/digestion|Digestion]] test
+
[[Team:Chiba/protocol/digestion|Digestion test]]
*[http://partsregistry.org/Part:BBa_R0079 BBa_R0079]①
*[http://partsregistry.org/Part:BBa_R0079 BBa_R0079]①
*[http://partsregistry.org/Part:BBa_R0071 BBa_R0071]②
*[http://partsregistry.org/Part:BBa_R0071 BBa_R0071]②
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</tr>
</tr>
</table>
</table>
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-
 
== 4,Septempber,2008 ==
== 4,Septempber,2008 ==
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[[Team:Chiba/jk/γ/Trl|TIME RESPONCE(liquid)]]
[[Team:Chiba/jk/γ/Trl|TIME RESPONCE(liquid)]]
-
[[Team:Chiba/protocol/digestion|Digestion]] test
+
[[Team:Chiba/protocol/digestion|Digestion test]]
*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2
*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2
*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]  x2
*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]  x2
Line 271: Line 270:
-->37℃ 30in
-->37℃ 30in
-
Gel Cheack
+
[[Team:Chiba/protocol/gelcheck|Gel Check]]
*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2①
*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2①
*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]  x2②
*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]  x2②

Latest revision as of 06:30, 24 October 2008

実験ログ

出力班

Contents

WEEK 3

31,August,2008

Digestion

  • [http://partsregistry.org/Part:BBa_R0010 BBa_R0010]①
  • PCR product([http://partsregistry.org/Part:BBa_J52008 BBa_J52008])②
Sample No. ①②
DNA 1010
SpeⅠ 10
PstⅠ 11
XbaⅠ 01
BSA(x100) 0.50.5
Buffer2 50
Buffer3 05
dH2O 28.528.5
TOTAL 5050

-->37℃ 3hour

-->①20μl refine

-->②20μl refine


1,September,2008

Ligation

  • vector:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]①
  • negative control:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]②
Sample No. ①②
vector 22
insert 30
Ligase Buffer 22
Ligase 11
dH2O 35
TOTAL 1010

-->R/T 2hour

Transformation

  • [http://partsregistry.org/Part:BBa_R0079 BBa_R0079](Las promoter)
  • [http://partsregistry.org/Part:BBa_R0071 BBa_R0071](RhlR promoter)
  • [http://partsregistry.org/Part:BBa_R0077 BBa_R0077](cinR promoter+RBS)
  • [http://partsregistry.org/Part:BBa_R0078 BBa_R0078](cinR promoter)
  • [http://partsregistry.org/Part:BBa_R0062 BBa_R0062](Lux promoter)


2,September,2008

TIME RESPONCE (Solid)

Colony PCR

  • [http://partsregistry.org/Part:BBa_R0010 BBa_R0010]+[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]
Sample No. ①
culture 1
Fwd primer 1.5
Rev primer 1.5
Thermo pol Buffer 3
dNTP mix 3
Taq DNA pol (NEB) 0.2 (1 unit)
dH2O 19.8
TOTAL 30

Mini prep

  • [http://partsregistry.org/Part:BBa_R0079 BBa_R0079]
  • [http://partsregistry.org/Part:BBa_R0071 BBa_R0071]
  • [http://partsregistry.org/Part:BBa_R0077 BBa_R0077]
  • [http://partsregistry.org/Part:BBa_R0078 BBa_R0078]
  • [http://partsregistry.org/Part:BBa_R0062 BBa_R0062]

3,September,2008

Digestion test

  • [http://partsregistry.org/Part:BBa_R0079 BBa_R0079]①
  • [http://partsregistry.org/Part:BBa_R0071 BBa_R0071]②
  • [http://partsregistry.org/Part:BBa_R0077 BBa_R0077]③
  • [http://partsregistry.org/Part:BBa_R0078 BBa_R0078]④
  • [http://partsregistry.org/Part:BBa_R0062 BBa_R0062]⑤
Sample No. ①~⑤
DNA tamplate 5
Buffer3 1
dH2O 4
EcoRⅠ 0.1
TOTAL 10

-->37℃30min

-->Gel check

Sample No. ①~⑤
DNA(digestion sample) 10
loading Dye 2
TOTAL 12

4,Septempber,2008

Transformation

  • Ligation product
  • rLUciferase([http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] ) x3
  • Venus YFP([http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]) x3

5,September,2008

TIME RESPONCE(liquid)

Digestion test

  • [http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2
  • [http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2
DNA(digested sample) 2
PstⅠ 0.1
BSA 0.8
Buffer3 1
dH2O 7
TOTAL 10

-->37℃ 30in

Gel Check

  • [http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2①
  • [http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2②
  • [http://partsregistry.org/Part:BBa_J52008 BBa_J52008](PCR Product)③
  • [http://partsregistry.org/Part:BBa_E2050 BBa_E2050]④
Sample No. ①~②③,④
DNA(digested sample) 101
loading Dye 21
TOTAL 126
  • Luciferase check-->no light


>Last Week

>Next Week


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