Team:Chiba/jk/β/week 4

From 2008.igem.org

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-->(6/9)
-->(6/9)
[[Team:Chiba/protocol/digestion|Digestion Test]]
[[Team:Chiba/protocol/digestion|Digestion Test]]
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::Sample DNA 1 : BBa_R0010(濃度未知)[http://partsregistry.org/Part:BBa_R0010]
+
::Sample DNA 1 : BBa_R0010(unknown)[http://partsregistry.org/Part:BBa_R0010]
::Sample DNA 2 : BBa_R0010(100ng/μL)[http://partsregistry.org/Part:BBa_R0010]
::Sample DNA 2 : BBa_R0010(100ng/μL)[http://partsregistry.org/Part:BBa_R0010]
Line 72: Line 72:
===8 September, 2008===
===8 September, 2008===
 +
-->(7/9)
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 +
'''[[Team:Chiba/protocol/ligation/gelex|Gel Extract]]'''
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 +
 +
:--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
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:::->15μL?
===9 September, 2008===
===9 September, 2008===
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-->(6/9)PCR of I9030
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 +
--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
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 +
--->'''[[Team:Chiba/protocol/ligation/gelex|Gel Extract]]'''
 +
 +
--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
===10 September, 2008===
===10 September, 2008===
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-->(10/9)
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:Colony Count
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#LuxI
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#LuxI with LVA
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#Las
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#Background(R0010)
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#Background(C0261)
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 +
Pick colony
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 +
'''[[Team:Chiba/protocol/PCR|Colony-PCR]]'''
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::Colony PCR of 10 colonies from ligation plate(9/9-(2)) and 3 colonies from ligation plate(9/9-(3)) one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
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:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<tr>
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<td width="257">DNA Template</td>
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<td>1</td>
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</tr>
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<tr>
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<td>dNTP mix</td>
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<td>5</td>
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</tr>
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<tr>
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<td>Foward Primer</td>
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<td>0.3</td>
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</tr>
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<tr>
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<td>Reverse Primer</td>
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<td>0.3</td>
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</tr>
 +
<tr>
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<td>taq DNA polymerase</td>
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<td>0.5</td>
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</tr>
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<tr>
 +
<td>Thermopol Buffer</td>
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<td>3</td>
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</tr>
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<tr>
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<td>Nuclease free water</td>
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<td>20</td>
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</tr>
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<tr>
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<td>TOTAL</td>
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<td>30μL</td>
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</tr>
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</table>
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 +
??
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:95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃
 +
??
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 +
-->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
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 +
--->(28/8)'''[[Team:Chiba/protocol/DNA Purification/sigma|Miniprep]]'''
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 +
elute 50μL
===11 September, 2008===
===11 September, 2008===
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</table>
</table>
-
===[[Team:Chiba/jk/β/weak 5|>next week]]===
+
 
 +
 
===[[Team:Chiba/jk/β/week 5|>next week]]===
===[[Team:Chiba/jk/β/week 5|>next week]]===

Latest revision as of 12:52, 30 September 2008

>送受信班

Contents

Week 4

>last week

7 September, 2008

-->(6/9) Digestion Test

Sample DNA 1 : BBa_R0010(unknown)[http://partsregistry.org/Part:BBa_R0010]
Sample DNA 2 : BBa_R0010(100ng/μL)[http://partsregistry.org/Part:BBa_R0010]
Sample No 1,2
Sample DNA 12
PstⅠ 0.5
SpeⅠ 0.5
Buffer 2 2
(×10)BSA 2
dH2O 3
TOTAL 20


--->Gel Check
Sample leftright
Sample DNA 55
Loading Dye 11
TOTAL 66

Left side:BBa_R0010 Sample No.1
Right side:BBa_R0010 Sample No.2

-->(8/9)

8 September, 2008

-->(7/9)

Gel Extract


--->Zymo Clean
->15μL?

9 September, 2008

-->(6/9)PCR of I9030

--->Gel Check

--->Gel Extract

--->Zymo Clean

10 September, 2008

-->(10/9)

Colony Count
  1. LuxI
  2. LuxI with LVA
  3. Las
  4. Background(R0010)
  5. Background(C0261)

Pick colony

Colony-PCR

Colony PCR of 10 colonies from ligation plate(9/9-(2)) and 3 colonies from ligation plate(9/9-(3)) one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
DNA Template 1
dNTP mix 5
Foward Primer 0.3
Reverse Primer 0.3
taq DNA polymerase 0.5
Thermopol Buffer 3
Nuclease free water 20
TOTAL 30μL

??

95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃

??

-->Gel Check

--->(28/8)Miniprep

elute 50μL

11 September, 2008

12 September, 2008

13 September, 2008

Gel Check
  1. BBa_C0161[http://partsregistry.org/Part:BBa_Co161](2007)
  2. BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
  3. BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
Chiba-0913.JPG
Sample No 1~3
Sample DNA 1
Loading Dye 1
dH2O 4
TOTAL 6
From left;


-->Digestion

  1. BBa_C0161[http://partsregistry.org/Part:BBa_Co161](2007)
  2. BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
  3. BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)


Sample No ①②③④
Sample DNA 5511
EcoRⅠ 0.5o.5--
SpeⅠ 0.50.5--
Buffer 1 11--
Buffer 3 --1-
BSA 11--
dH2O 227.5-
TOTAL 1010101


>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%B2/week_4"