Team:Chiba/Calendar-Home/25 August 2008

From 2008.igem.org

(Difference between revisions)
(New page: '''OUTPUT TEAM''' <br>digestion <br>Lux1 <br>LUx2(１と２は別のコロニーをつついた） <br>Craig YFP <br>DNA 3μl <br>Buffer(x10) 1μl <br>dH2O 　6μl <br>EcoRⅠ 0.5μl <b...)
(→Team:Output)
 
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'''OUTPUT TEAM'''
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>[[Team:Chiba|Home]] | [[Team:Chiba/Notebook|Notebook]]
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<br>digestion
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-
<br>Lux1
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<br>LUx2(１と２は別のコロニーをつついた）
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-
<br>Craig YFP
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<br>DNA  3μl
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[[Team:Chiba/Calendar-Home/24 August 2008|24 August 2008 <]]|[[Team:Chiba/Calendar-Home/26 August 2008|> 26 August 2008]]
-
<br>Buffer(x10) 1μl
+
 
-
<br>dH2O 　6μl
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==Laboratory work==
-
<br>EcoRⅠ 0.5μl
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===Team:Input===
-
<br>---------------
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The following glycerol Stock was made.
-
<br>10μl
+
 
-
<br> 1hour 37℃
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[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](insert check OK)
-
<br>ゲル電(135v 30min)
+
 
-
<br>もとのDNA  1μl           
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<BR>Picked the colony and incubate with 2mL of LB-Ampicillin Medium for 12 hours at 37 degrees.
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<br>loading dye(x6) 1μl     
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Took 180&mu;l from culture,and mixed with 880&mu;lLB-Amp Medium.(OD=0.485)
-
<br>dH2O 3μl               
+
 
-
<br>--------------------
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<table width="210" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
-
<br>     6μl                 
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<td width="257">Liquid culture(OD=0.495)</td>
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<br> 切ったDNA　　10μl
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<td>0.67μl</td>
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<br> loading dye(x6) 2μl 
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<tr>
-
<br>----------------------
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<td>60%Glycerol</td>
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<br>     12μl
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<td>0.33μl</td>
 +
</tr>
 +
<tr>
 +
<td>20%Glycerol Stock</td>
 +
<td>1.00ml</td>
 +
</tr>
 +
</table>
 +
 
 +
===Team:Communication===
 +
:(24/8)--->'''[[Team:Chiba/protocol/digestion|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)
 +
 
 +
<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>
 +
</tr>
 +
<tr>
 +
<td>Sample DNA(&mu;L)</td>
 +
<td>5</td><td>5</td><td>1</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>EcoRI(&mu;L)</td>
 +
<td>0.5</td><td>0.5</td><td>-</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>SpeI(&mu;L)</td>
 +
<td>0.5</td><td>0.5</td><td>-</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 1</td>
 +
<td>1</td><td>1</td><td>-</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 3(&mu;L)</td>
 +
<td>-</td><td>-</td><td>1</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>BSA(&mu;L)</td>
 +
<td>1</td><td>1</td><td>-</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<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(&mu;L)</td>
 +
<td>10</td><td>10</td><td>10</td><td>1</td>
 +
</tr>
 +
</table>
 +
 
 +
:--->[[Team:Chiba/protocol/gelcheck|Gel Check]]
 +
{|align="justify"
 +
|[[Image:Chiba-0825-1.JPG]]
 +
|
 +
:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample No.</td>
 +
<td>1~3</td><td>4</td>
 +
</tr>
 +
<tr>
 +
<td>Sample DNA(&mu;L)</td>
 +
<td>10</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>Loading Dye(&mu;L)</td>
 +
<td>2</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O(&mu;L)</td>
 +
<td>-</td><td>4</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL(&mu;L)</td>
 +
<td>12</td><td>6</td>
 +
</tr>
 +
</table>
 +
: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)
 +
|}
 +
 
 +
 
 +
--->[[Team:Chiba/protocol/digestion|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)
 +
 
 +
<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>
 +
</tr>
 +
<tr>
 +
<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(&mu;L)</td>
 +
<td>2</td><td>2</td><td>2</td>
 +
</tr>
 +
<tr>
 +
<td>XbaI(&mu;L)</td>
 +
<td>2</td><td>2</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>SpeI(&mu;L)</td>
 +
<td>-</td><td>-</td><td>2</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 2(&mu;L)</td>
 +
<td>-</td><td>-</td><td>5</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 3(&mu;L)</td>
 +
<td>10</td><td>10</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>BSA(&mu;L)</td>
 +
<td>10</td><td>10</td><td>5</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O(&mu;L)</td>
 +
<td>16</td><td>16</td><td>16</td>
 +
</tr>
 +
<tr>
 +
<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_J33202 BBa_J33202](2)
 +
*BL21(3)
 +
*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](4)
 +
*[http://partsregistry.org/Part:BBa_I712019 BBa_I712019](5)
 +
 
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">No.</td>
 +
<td>1</td><td>2</td><td>3</td><td>4</td><td>5</td>
 +
</tr>
 +
<tr>
 +
<td>DNA tamplate(&mu;L)</td>
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<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<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(&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(&mu;L)</td>
 +
<td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
 +
</tr>
 +
<tr>
 +
<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)(&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(&mu;L)</td>
 +
<td>68</td><td>68</td><td>68</td><td>68</td><td>68</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL(&mu;L)</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="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">No.</td>
 +
<td>1</td><td>2</td><td>3</td><td>4</td><td>5</td>
 +
</tr>
 +
<tr>
 +
<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(&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(&mu;L)</td>
 +
<td>4</td><td>4</td><td>4</td><td>4</td><td>4</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL(&mu;L)</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]]
 +
*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
 +
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
 +
 
 +
 
 +
[[Team:Chiba/protocol/transformation|Transformation]]
 +
*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
 +
*PUC19
 +
*pGFPUV
 +
*pGEX Venus YFP
 +
*Membren Venus YFP
 +
[[Team:Chiba/protocol/PCR|PCR]]
 +
*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001](1)
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">No.</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>DNA tamplate(&mu;L)</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>FW primer(Venus)(&mu;L)</td>
 +
<td>5</td>
 +
</tr>
 +
<tr>
 +
<td>VR primer(&mu;L)</td>
 +
<td>5</td>
 +
</tr>
 +
<tr>
 +
<td>dNTPmix(&mu;L)</td>
 +
<td>10</td>
 +
</tr>
 +
<tr>
 +
<td>Thermo pol buffer(&mu;L)</td>
 +
<td>10</td>
 +
</tr>
 +
<tr>
 +
<td>DNA pol(VENT)(&mu;L)</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O(&mu;L)</td>
 +
<td>68</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL(&mu;L)</td>
 +
<td>100</td>
 +
</tr>
 +
</table>

Latest revision as of 06:15, 30 October 2008

>Home | Notebook

24 August 2008 <|> 26 August 2008

Contents

Laboratory work

Team:Input

The following glycerol Stock was made.

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
  1. Double Digestion:BBa_J04500(2007)
  2. Double Digesiton:BBa_R0010(2007)
  3. Single Digestion:BBa_R0010(2007)
  4. BBa_R0010(2007)
Sample No. 1234
Sample DNA(μL) 5511
EcoRI(μL) 0.50.5--
SpeI(μL) 0.50.5--
Buffer 1 11--
Buffer 3(μL) --1-
BSA(μL) 11--
dH2O(μL) 227.5-
TOTAL(μL) 1010101
--->Gel Check
Chiba-0825-1.JPG
Sample No. 1~34
Sample DNA(μL) 101
Loading Dye(μL) 21
dH2O(μL) -4
TOTAL(μL) 126
From right;
  1. Double Digestion:BBa_J04500(2007) -> OK
  2. Double Digesiton:BBa_R0010(2007) -> ?(low)
  3. Single Digestion:BBa_R0010(2007) -> ?(low)
  4. BBa_R0010(2007) -> OK(low)


--->Digestion (Double Digestion)

  1. BBa_C0178
  2. BBa_C0170
  3. BBa_J04500(2007)
Sample No. 123
Sample DNA(μL) 33ng/μl×60μl33ng/μl×60μl100ng/μl×20μl
PstI(μL) 222
XbaI(μL) 22-
SpeI(μL) --2
Buffer 2(μL) --5
Buffer 3(μL) 1010-
BSA(μL) 10105
dH2O(μL) 161616
TOTAL(μL) 10010052

Team:Output

PCR

No. 12345
DNA tamplate(μL) 11111
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) 1010101010
Thermo pol buffer(μL) 1010101010
DNA pol(VENT)(μL) 11111
dH2O(μL) 6868686868
TOTAL(μL) 100100100100100

-->Gel Check


No. 12345
DNA tamplate(μL) 11111
loading Dye(μL) 11111
dH2O(μL) 44444
TOTAL(μL) 66666

Mini prep


Transformation

  • BBa_E2030
  • PUC19
  • pGFPUV
  • pGEX Venus YFP
  • Membren Venus YFP

PCR

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
Retrieved from "http://2008.igem.org/Team:Chiba/Calendar-Home/25_August_2008"