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...)
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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/25 August 2008|25 August 2008 <]]|[[Team:Chiba/Calendar-Home/26 August 2008|> 26 August 2008]]
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<br>Buffer(x10) 1μl
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<br>dH2O 　6μl
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==Laboratory work==
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<br>EcoRⅠ 0.5μl
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===Team:Input===
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<br>---------------
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[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](insert check済み)のグリセロールストックづくり
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<br>10μl
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<BR><BR>プレートに生えたコロニーをpickして、試験管で2ml培養(LB-Amp,37℃,12h)
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<br> 1hour 37℃
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<BR>培養した試験管から180μl,LB-Ampを880μlとって、OD=0.495にした。
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<br>ゲル電(135v 30min)
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<BR><table width="210" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<br>もとのDNA  1μl           
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<td width="257">OD=0.495培養液</td>
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<br>loading dye(x6) 1μl     
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<td>0.67μl</td>
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<br>dH2O 3μl               
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<tr>
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<br>--------------------
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<td>60%グリセロール</td>
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<br>     6μl                 
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<td>0.33μl</td>
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<br> 切ったDNA　　10μl
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</tr>
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<br> loading dye(x6)  2μl 
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<tr>
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<br>----------------------
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<td>20%グリセロールストック(計)</td>
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<br>     12μl
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<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</td>
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<td>5</td><td>5</td><td>1</td><td>1</td>
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</tr>
 +
<tr>
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<td>EcoRⅠ</td>
 +
<td>0.5</td><td>0.5</td><td>-</td><td>-</td>
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</tr>
 +
<tr>
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<td>SpeⅠ</td>
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<td>0.5</td><td>0.5</td><td>-</td><td>-</td>
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</tr>
 +
<tr>
 +
<td>Buffer 1</td>
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<td>1</td><td>1</td><td>-</td><td>-</td>
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</tr>
 +
<tr>
 +
<td>Buffer 3</td>
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<td>-</td><td>-</td><td>1</td><td>-</td>
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</tr>
 +
<tr>
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<td>BSA</td>
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<td>1</td><td>1</td><td>-</td><td>-</td>
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</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
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<td>2</td><td>2</td><td>7.5</td><td>-</td>
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</tr>
 +
<tr>
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<td>TOTAL</td>
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<td>10</td><td>10</td><td>10</td><td>1</td>
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</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>
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<td>1~3</td><td>4</td>
 +
</tr>
 +
<tr>
 +
<td>Sample DNA</td>
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<td>10</td><td>1</td>
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</tr>
 +
<tr>
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<td>Loading Dye</td>
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<td>2</td><td>1</td>
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</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
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<td>-</td><td>4</td>
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</tr>
 +
<tr>
 +
<td>TOTAL</td>
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<td>12</td><td>6</td>
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</tr>
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</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>
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<td width="257">Sample No.</td>
 +
<td>1</td><td>2</td><td>3</td>
 +
</tr>
 +
<tr>
 +
<td>Sample DNA</td>
 +
<td>33ng/μl×60μl</td><td>33ng/μl×60μl</td><td>100ng/μl×20μl</td>
 +
</tr>
 +
<tr>
 +
<td>PstⅠ</td>
 +
<td>2</td><td>2</td><td>2</td>
 +
</tr>
 +
<tr>
 +
<td>XbaⅠ</td>
 +
<td>2</td><td>2</td><td>-</td>
 +
</tr>
 +
<tr>
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<td>SpeⅠ</td>
 +
<td>-</td><td>-</td><td>2</td>
 +
</tr>
 +
<tr>
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<td>Buffer 2</td>
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<td>-</td><td>-</td><td>5</td>
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</tr>
 +
<tr>
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<td>Buffer 3</td>
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<td>10</td><td>10</td><td>-</td>
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</tr>
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<tr>
 +
<td>BSA</td>
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<td>10</td><td>10</td><td>5</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
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<td>16</td><td>16</td><td>16</td>
 +
</tr>
 +
<tr>
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<td>TOTAL</td>
 +
<td>100</td><td>100</td><td>52</td>
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</tr>
 +
</table>
 +
 
 +
 
 +
 
 +
===Team:Output===[[Team:Chiba/protocol/PCR|PCR]]
 +
*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]①
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*[http://partsregistry.org/Part:BBa_J33202 BBa_J33202]②
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*BL21③
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*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]④
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*[http://partsregistry.org/Part:BBa_I712019 BBa_I712019]⑤
 +
 
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<tr>
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<td width="257">Sample No.</td>
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<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
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</tr>
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<tr>
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<td>DNA tamplate</td>
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<td>1</td><td>1</td><td>1</td><td>1</td>><td>1</td>
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</tr>
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<tr>
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<td>FW primer</td>
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<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>
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<tr>
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<td>VR primer</td>
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<td>5(VR)</td><td>5(LacZ_rev)</td><td>5(LacZ_rev)</td><td>5(VR)</td><td>5(VR)</td>
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</tr>
 +
<tr>
 +
<td>dNTPmix</td>
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<td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
 +
</tr>
 +
<tr>
 +
<td>Thermo pol buffer</td>
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<td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
 +
</tr>
 +
<tr>
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<td>DNA pol(VENT)</td>
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<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
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</tr>
 +
<tr>
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<td>dH<sub>2</sub>O</td>
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<td>68</td><td>68</td><td>68</td><td>68</td><td>68</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
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<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">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>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>
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<td>TOTAL</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]]
 +
*Venus
 +
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
 +
 
 +
 
 +
[[Team:Chiba/protocol/transformation|Transformation]]
 +
*Venus
 +
*PUC19
 +
*pGFPUV
 +
*pGEX Venus YFP
 +
*Membren Venus YFP
 +
[[Team:Chiba/protocol/PCR|PCR]]
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*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001]①
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample No.</td>
 +
<td>①</td>
 +
</tr>
 +
<tr>
 +
<td>DNA tamplate</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>FW primer(Venus)</td>
 +
<td>5</td>
 +
</tr>
 +
<tr>
 +
<td>VR primer</td>
 +
<td>5</td>
 +
</tr>
 +
<tr>
 +
<td>dNTPmix</td>
 +
<td>10</td>
 +
</tr>
 +
<tr>
 +
<td>Thermo pol buffer</td>
 +
<td>10</td>
 +
</tr>
 +
<tr>
 +
<td>DNA pol(VENT)</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
 +
<td>68</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>100</td>
 +
</tr>
 +
</table>

Revision as of 07:05, 24 October 2008

>Home | Notebook

25 August 2008 <|> 26 August 2008

Laboratory work

Team:Input

BBa_J22136(insert check済み)のグリセロールストックづくり

プレートに生えたコロニーをpickして、試験管で2ml培養(LB-Amp,37℃,12h)
培養した試験管から180μl,LB-Ampを880μlとって、OD=0.495にした。


OD=0.495培養液 0.67μl
60%グリセロール 0.33μl
20%グリセロールストック(計) 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 5511
EcoRⅠ 0.50.5--
SpeⅠ 0.50.5--
Buffer 1 11--
Buffer 3 --1-
BSA 11--
dH2O 227.5-
TOTAL 1010101
--->Gel Check
Chiba-0825-1.JPG
Sample No. 1~34
Sample DNA 101
Loading Dye 21
dH2O -4
TOTAL 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 33ng/μl×60μl33ng/μl×60μl100ng/μl×20μl
PstⅠ 222
XbaⅠ 22-
SpeⅠ --2
Buffer 2 --5
Buffer 3 1010-
BSA 10105
dH2O 161616
TOTAL 10010052


===Team:Output===PCR

>
Sample No. ①②③④⑤
DNA tamplate 11111
FW primer 5(Venus_YFP_fwd)5(LacZ_fwd)5(LacZ_fwd)5(rLUC_fwd)5(fLuc_fwd)
VR primer 5(VR)5(LacZ_rev)5(LacZ_rev)5(VR)5(VR)
dNTPmix 1010101010
Thermo pol buffer 1010101010
DNA pol(VENT) 11111
dH2O 6868686868
TOTAL 100100100100100

-->Gel Check


>
Sample No. ①②③④⑤
DNA tamplate 11111
loading Dye 11111
dH2O 44444
TOTAL 66666

Mini prep


Transformation

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

PCR

Sample No. ①
DNA tamplate 1
FW primer(Venus) 5
VR primer 5
dNTPmix 10
Thermo pol buffer 10
DNA pol(VENT) 1
dH2O 68
TOTAL 100
Retrieved from "http://2008.igem.org/Team:Chiba/Calendar-Home/25_August_2008"