Team:Chiba/jk/γ/week 3

From 2008.igem.org

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-
<br>
 
[[Team:Chiba/jk|実験ログ]]
[[Team:Chiba/jk|実験ログ]]
-
<br>[[Team:Chiba/jk/γ|出力班]]
+
 
 +
[[Team:Chiba/jk/γ|出力班]]
== WEEK 3 ==
== WEEK 3 ==
Line 9: Line 9:
== 31,August,2008 ==
== 31,August,2008 ==
-
<br>digestion
+
[[Team:Chiba/protocol/digestion|Digestion]]
-
<br>BBa_R0010①
+
 
-
<br>PCR product(BBa_J52008)②
+
*[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]①
 +
*PCR product([http://partsregistry.org/Part:BBa_J52008 BBa_J52008])②
 +
 
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<tr>
<tr>
-
<td width="257">Sample No</td>
+
<td width="257">Sample No.</td>
<td>①</td><td>②</td>
<td>①</td><td>②</td>
</tr>
</tr>
Line 46: Line 48:
</tr>
</tr>
<tr>
<tr>
-
<td>dH20</td>
+
<td>dH<sub>2</sub>O</td>
<td>28.5</td><td>28.5</td>
<td>28.5</td><td>28.5</td>
</tr>
</tr>
Line 54: Line 56:
</tr>
</tr>
</table>
</table>
-
<br>->37℃ 3hour
+
 
-
<br>->①20μl refine
+
-->37℃ 3hour
-
<br>->②20μl refine
+
 
 +
-->①20μl refine
 +
 
 +
-->②20μl refine
== 1,September,2008 ==
== 1,September,2008 ==
-
<br>Ligation
+
[[Team:Chiba/protocol/ligation/ligation|Ligation]]
-
<br>vector:BBa_R0010 insert:BBa_J52008①
+
*vector:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]①
-
<br>negative control:BBa_R0010②
+
*negative control:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]②
 +
 
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<tr>
<tr>
-
<td width="257">Sample No</td>
+
<td width="257">Sample No.</td>
<td>①</td><td>②</td>
<td>①</td><td>②</td>
</tr>
</tr>
Line 86: Line 92:
</tr>
</tr>
<tr>
<tr>
-
<td>dH2O</td>
+
<td>dH<sub>2</sub>O</td>
<td>3</td><td>5</td>
<td>3</td><td>5</td>
</tr>
</tr>
 +
<tr>
<td>TOTAL</td>
<td>TOTAL</td>
<td>10</td><td>10</td>
<td>10</td><td>10</td>
</tr>
</tr>
</table>
</table>
-
<br>->R/T 2hour
 
-
<br>TRANSFORMATION
+
-->R/T 2hour
-
<br>BBa_R0079(Las promoter)
+
-
<br>BBa_R0071(RhlR promoter)
+
-
<br>BBa_R0077(cinR promoter+RBS)
+
-
<br>BBa_R0078(cnR promoter)
+
-
<br>BBa_R0062(LUX promoter)
+
 +
[[Team:Chiba/protocol/transformation|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)
Line 107: Line 115:
== 2,September,2008 ==
== 2,September,2008 ==
-
<br>[[Team:Chiba/jk/γ/trs|TIME RESPONCE (Solid)]]
+
[[Team:Chiba/jk/γ/trs|TIME RESPONCE (Solid)]]
 +
 
 +
Colony PCR
 +
*[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]+[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]
-
<br>COLONY PCR
 
-
<br>BBa_R0010+BBa_J52008
 
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<tr>
<tr>
-
<td width="257">Sample No</td>
+
<td width="257">Sample No.</td>
<td>①</td>
<td>①</td>
</tr>
</tr>
Line 120: Line 129:
<td>1</td>
<td>1</td>
</tr>
</tr>
 +
<tr>
<td>Fwd primer</td>
<td>Fwd primer</td>
<td>1.5</td>
<td>1.5</td>
Line 140: Line 150:
</tr>
</tr>
<tr>
<tr>
-
<td>dH2O</td>
+
<td>dH<sub>2</sub>O</td>
<td>19.8</td>
<td>19.8</td>
</tr>
</tr>
 +
<tr>
<td>TOTAL</td>
<td>TOTAL</td>
<td>30</td>
<td>30</td>
</tr>
</tr>
</table>
</table>
-
<br>MINI Prep
+
 
-
<br>BBa_R0079
+
[[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]]
-
<br>BBa_R0071
+
*[http://partsregistry.org/Part:BBa_R0079 BBa_R0079]
-
<br>BBa_R0077
+
*[http://partsregistry.org/Part:BBa_R0071 BBa_R0071]
-
<br>BBa_R0078
+
*[http://partsregistry.org/Part:BBa_R0077 BBa_R0077]
-
<br>BBa_R0062
+
*[http://partsregistry.org/Part:BBa_R0078 BBa_R0078]
 +
*[http://partsregistry.org/Part:BBa_R0062 BBa_R0062]
== 3,September,2008 ==
== 3,September,2008 ==
-
<br>Digestion test
+
[[Team:Chiba/protocol/digestion|Digestion test]]
-
<br>BBa_R0079①
+
*[http://partsregistry.org/Part:BBa_R0079 BBa_R0079]①
-
<br>BBa_R0071②
+
*[http://partsregistry.org/Part:BBa_R0071 BBa_R0071]②
-
<br>BBa_R0077③
+
*[http://partsregistry.org/Part:BBa_R0077 BBa_R0077]③
-
<br>BBa_R0078④
+
*[http://partsregistry.org/Part:BBa_R0078 BBa_R0078]④
-
<br>BBa_R0062⑤
+
*[http://partsregistry.org/Part:BBa_R0062 BBa_R0062]⑤
 +
 
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<tr>
<tr>
-
<td width="257">Sample No</td>
+
<td width="257">Sample No.</td>
<td>①~⑤</td>
<td>①~⑤</td>
</tr>
</tr>
Line 176: Line 189:
</tr>
</tr>
<tr>
<tr>
-
<td>dH2O</td>
+
<td>dH<sub>2</sub>O</td>
<td>4</td>
<td>4</td>
</tr>
</tr>
Line 183: Line 196:
<td>0.1</td>
<td>0.1</td>
</tr>
</tr>
 +
<tr>
<td>TOTAL</td>
<td>TOTAL</td>
<td>10</td>
<td>10</td>
</tr>
</tr>
</table>
</table>
-
<br>->37℃30min
+
 
-
<br>->Gel cheack
+
-->37℃30min
 +
 
 +
-->[[Team:Chiba/protocol/gelcheck|Gel check]]
 +
 
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<tr>
<tr>
-
<td width="257">Sample No</td>
+
<td width="257">Sample No.</td>
<td>①~⑤</td>
<td>①~⑤</td>
</tr>
</tr>
Line 207: Line 224:
</tr>
</tr>
</table>
</table>
-
 
-
 
== 4,Septempber,2008 ==
== 4,Septempber,2008 ==
 +
[[Team:Chiba/protocol/transformation|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 ==
 +
[[Team:Chiba/jk/γ/Trl|TIME RESPONCE(liquid)]]
-
== 5,September,2008 ==
+
[[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_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]  x2
-
<br>[[Team:Chiba/jk/γ/Trl|TIME RESPONCE(liquid)]]
 
-
<br>Digestion test
 
-
<br>BBa_J52008+BBa_F2620
 
-
<br>BBa_E2030+BBa_F2620
 
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<tr>
<tr>
-
<td width="257">Sample No</td>
+
<td width="257">DNA(digested sample)</td>
-
<td>each</td>
+
<td>2</td>
</tr>
</tr>
<tr>
<tr>
-
<td>DNA(digestion sample)</td>
+
<td>PstⅠ</td>
 +
<td>0.1</td>
 +
</tr>
 +
<tr>
 +
<td>BSA</td>
 +
<td>0.8</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer3</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
 +
<td>7</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
<td>10</td>
<td>10</td>
 +
</tr>
 +
</table>
 +
 +
-->37℃ 30in
 +
 +
[[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_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
 +
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample No.</td>
 +
<td>①~②</td><td>③,④</td>
 +
</tr>
 +
<tr>
 +
<td>DNA(digested sample)</td>
 +
<td>10</td><td>1</td>
</tr>
</tr>
<tr>
<tr>
<td>loading Dye</td>
<td>loading Dye</td>
-
<td>2</td>
+
<td>2</td><td>1</td>
</tr>
</tr>
<tr>
<tr>
<td>TOTAL</td>
<td>TOTAL</td>
-
<td>12</td>
+
<td>12</td><td>6</td>
</tr>
</tr>
</table>
</table>
-
<br>Luciferase cheack->no light
+
*Luciferase check-->no light
 +
 
 +
[[Team:Chiba/jk/γ/week 2|>Last Week]]
-
<br>[[Team:Chiba/jk/γ/week 2|>Last Week]]
+
[[Team:Chiba/jk/γ/week 4|>Next Week]]
-
<br>[[Team:Chiba/jk/γ/week 4|>Next Week]]
+

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


ホーム メンバー紹介 プロジェクト紹介 Parts Submitted to the Registry モデリング ノート
Retrieved from "http://2008.igem.org/Team:Chiba/jk/%CE%B3/week_3"