Team:Chiba/Calendar-Home/25 August 2008

From 2008.igem.org

(Difference between revisions)
(Team:Input)
(Team:Output)
 
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>[[Team:Chiba|Home]] | [[Team:Chiba/Notebook|Notebook]]
>[[Team:Chiba|Home]] | [[Team:Chiba/Notebook|Notebook]]
-
[[Team:Chiba/Calendar-Home/25 August 2008|24 August 2008 <]]|[[Team:Chiba/Calendar-Home/26 August 2008|> 26 August 2008]]
+
[[Team:Chiba/Calendar-Home/24 August 2008|24 August 2008 <]]|[[Team:Chiba/Calendar-Home/26 August 2008|> 26 August 2008]]
==Laboratory work==
==Laboratory work==
===Team:Input===
===Team:Input===
-
[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
+
The following glycerol Stock was made.
-
<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)
+
 
-
<BR>培養した試験管から180μl,LB-Ampを880μlとって、OD=0.495にした。
+
[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](insert check OK)
-
<BR><table width="210" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
 
-
<td width="257">OD=0.495培養液</td>
+
<BR>Picked the colony and incubate with 2mL of LB-Ampicillin Medium for 12 hours at 37 degrees.
 +
Took 180&mu;l from culture,and mixed with 880&mu;lLB-Amp Medium.(OD=0.485)
 +
 
 +
<table width="210" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<td width="257">Liquid culture(OD=0.495)</td>
<td>0.67μl</td>
<td>0.67μl</td>
<tr>
<tr>
-
<td>60%グリセロール</td>
+
<td>60%Glycerol</td>
<td>0.33μl</td>
<td>0.33μl</td>
</tr>
</tr>
<tr>
<tr>
-
<td>20%グリセロールストック(計)</td>
+
<td>20%Glycerol Stock</td>
<td>1.00ml</td>
<td>1.00ml</td>
</tr>
</tr>
</table>
</table>
-
 
===Team:Communication===
===Team:Communication===
Line 35: Line 38:
</tr>
</tr>
<tr>
<tr>
-
<td>Sample DNA</td>
+
<td>Sample DNA(&mu;L)</td>
<td>5</td><td>5</td><td>1</td><td>1</td>
<td>5</td><td>5</td><td>1</td><td>1</td>
</tr>
</tr>
<tr>
<tr>
-
<td>EcoRⅠ</td>
+
<td>EcoRI(&mu;L)</td>
<td>0.5</td><td>0.5</td><td>-</td><td>-</td>
<td>0.5</td><td>0.5</td><td>-</td><td>-</td>
</tr>
</tr>
<tr>
<tr>
-
<td>SpeⅠ</td>
+
<td>SpeI(&mu;L)</td>
<td>0.5</td><td>0.5</td><td>-</td><td>-</td>
<td>0.5</td><td>0.5</td><td>-</td><td>-</td>
</tr>
</tr>
Line 51: Line 54:
</tr>
</tr>
<tr>
<tr>
-
<td>Buffer 3</td>
+
<td>Buffer 3(&mu;L)</td>
<td>-</td><td>-</td><td>1</td><td>-</td>
<td>-</td><td>-</td><td>1</td><td>-</td>
</tr>
</tr>
<tr>
<tr>
-
<td>BSA</td>
+
<td>BSA(&mu;L)</td>
<td>1</td><td>1</td><td>-</td><td>-</td>
<td>1</td><td>1</td><td>-</td><td>-</td>
</tr>
</tr>
<tr>
<tr>
-
<td>dH<sub>2</sub>O</td>
+
<td>dH<sub>2</sub>O(&mu;L)</td>
<td>2</td><td>2</td><td>7.5</td><td>-</td>
<td>2</td><td>2</td><td>7.5</td><td>-</td>
</tr>
</tr>
<tr>
<tr>
-
<td>TOTAL</td>
+
<td>TOTAL(&mu;L)</td>
<td>10</td><td>10</td><td>10</td><td>1</td>
<td>10</td><td>10</td><td>10</td><td>1</td>
</tr>
</tr>
Line 78: Line 81:
</tr>
</tr>
<tr>
<tr>
-
<td>Sample DNA</td>
+
<td>Sample DNA(&mu;L)</td>
<td>10</td><td>1</td>
<td>10</td><td>1</td>
</tr>
</tr>
<tr>
<tr>
-
<td>Loading Dye</td>
+
<td>Loading Dye(&mu;L)</td>
<td>2</td><td>1</td>
<td>2</td><td>1</td>
</tr>
</tr>
<tr>
<tr>
-
<td>dH<sub>2</sub>O</td>
+
<td>dH<sub>2</sub>O(&mu;L)</td>
<td>-</td><td>4</td>
<td>-</td><td>4</td>
</tr>
</tr>
<tr>
<tr>
-
<td>TOTAL</td>
+
<td>TOTAL(&mu;L)</td>
<td>12</td><td>6</td>
<td>12</td><td>6</td>
</tr>
</tr>
Line 114: Line 117:
</tr>
</tr>
<tr>
<tr>
-
<td>Sample DNA</td>
+
<td>Sample DNA(&mu;L)</td>
<td>33ng/μl×60μl</td><td>33ng/μl×60μl</td><td>100ng/μl×20μl</td>
<td>33ng/μl×60μl</td><td>33ng/μl×60μl</td><td>100ng/μl×20μl</td>
</tr>
</tr>
<tr>
<tr>
-
<td>PstⅠ</td>
+
<td>PstI(&mu;L)</td>
<td>2</td><td>2</td><td>2</td>
<td>2</td><td>2</td><td>2</td>
</tr>
</tr>
<tr>
<tr>
-
<td>XbaⅠ</td>
+
<td>XbaI(&mu;L)</td>
<td>2</td><td>2</td><td>-</td>
<td>2</td><td>2</td><td>-</td>
</tr>
</tr>
<tr>
<tr>
-
<td>SpeⅠ</td>
+
<td>SpeI(&mu;L)</td>
<td>-</td><td>-</td><td>2</td>
<td>-</td><td>-</td><td>2</td>
</tr>
</tr>
<tr>
<tr>
-
<td>Buffer 2</td>
+
<td>Buffer 2(&mu;L)</td>
<td>-</td><td>-</td><td>5</td>
<td>-</td><td>-</td><td>5</td>
</tr>
</tr>
<tr>
<tr>
-
<td>Buffer 3</td>
+
<td>Buffer 3(&mu;L)</td>
<td>10</td><td>10</td><td>-</td>
<td>10</td><td>10</td><td>-</td>
</tr>
</tr>
<tr>
<tr>
-
<td>BSA</td>
+
<td>BSA(&mu;L)</td>
<td>10</td><td>10</td><td>5</td>
<td>10</td><td>10</td><td>5</td>
</tr>
</tr>
<tr>
<tr>
-
<td>dH<sub>2</sub>O</td>
+
<td>dH<sub>2</sub>O(&mu;L)</td>
<td>16</td><td>16</td><td>16</td>
<td>16</td><td>16</td><td>16</td>
</tr>
</tr>
<tr>
<tr>
-
<td>TOTAL</td>
+
<td>TOTAL(&mu;L)</td>
<td>100</td><td>100</td><td>52</td>
<td>100</td><td>100</td><td>52</td>
</tr>
</tr>
</table>
</table>
-
 
-
 
===Team:Output===
===Team:Output===
[[Team:Chiba/protocol/PCR|PCR]]
[[Team:Chiba/protocol/PCR|PCR]]
-
*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
+
*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030](1)
-
*[http://partsregistry.org/Part:BBa_J33202 BBa_J33202]
+
*[http://partsregistry.org/Part:BBa_J33202 BBa_J33202](2)
-
*BL21③
+
*BL21(3)
-
*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]
+
*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](4)
-
*[http://partsregistry.org/Part:BBa_I712019 BBa_I712019]
+
*[http://partsregistry.org/Part:BBa_I712019 BBa_I712019](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">No.</td>
-
<td></td><td></td><td></td><td></td><td></td>
+
<td>1</td><td>2</td><td>3</td><td>4</td><td>5</td>
</tr>
</tr>
<tr>
<tr>
-
<td>DNA tamplate</td>
+
<td>DNA tamplate(&mu;L)</td>
-
<td>1</td><td>1</td><td>1</td><td>1</td>><td>1</td>
+
<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
</tr>
</tr>
<tr>
<tr>
-
<td>FW primer</td>
+
<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>
<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>
<tr>
<tr>
-
<td>VR primer</td>
+
<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>
<td>5(VR)</td><td>5(LacZ_rev)</td><td>5(LacZ_rev)</td><td>5(VR)</td><td>5(VR)</td>
</tr>
</tr>
<tr>
<tr>
-
<td>dNTPmix</td>
+
<td>dNTPmix(&mu;L)</td>
<td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
<td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
</tr>
</tr>
<tr>
<tr>
-
<td>Thermo pol buffer</td>
+
<td>Thermo pol buffer(&mu;L)</td>
<td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
<td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
</tr>
</tr>
<tr>
<tr>
-
<td>DNA pol(VENT)</td>
+
<td>DNA pol(VENT)(&mu;L)</td>
<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
</tr>
</tr>
<tr>
<tr>
-
<td>dH<sub>2</sub>O</td>
+
<td>dH<sub>2</sub>O(&mu;L)</td>
<td>68</td><td>68</td><td>68</td><td>68</td><td>68</td>
<td>68</td><td>68</td><td>68</td><td>68</td><td>68</td>
</tr>
</tr>
<tr>
<tr>
-
<td>TOTAL</td>
+
<td>TOTAL(&mu;L)</td>
<td>100</td><td>100</td><td>100</td><td>100</td><td>100</td>
<td>100</td><td>100</td><td>100</td><td>100</td><td>100</td>
</tr>
</tr>
Line 205: Line 206:
<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">No.</td>
-
<td></td><td></td><td></td><td></td><td></td>
+
<td>1</td><td>2</td><td>3</td><td>4</td><td>5</td>
</tr>
</tr>
<tr>
<tr>
-
<td>DNA tamplate</td>
+
<td>DNA tamplate(&mu;L)</td>
-
<td>1</td><td>1</td><td>1</td><td>1</td>><td>1</td>
+
<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
</tr>
</tr>
<tr>
<tr>
-
<td>loading Dye</td>
+
<td>loading Dye(&mu;L)</td>
<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
</tr>
</tr>
<tr>
<tr>
-
<td>dH<sub>2</sub>O</td>
+
<td>dH<sub>2</sub>O(&mu;L)</td>
<td>4</td><td>4</td><td>4</td><td>4</td><td>4</td>
<td>4</td><td>4</td><td>4</td><td>4</td><td>4</td>
</tr>
</tr>
<tr>
<tr>
-
<td>TOTAL</td>
+
<td>TOTAL(&mu;L)</td>
<td>6</td><td>6</td><td>6</td><td>6</td><td>6</td>
<td>6</td><td>6</td><td>6</td><td>6</td><td>6</td>
</tr>
</tr>
Line 227: Line 228:
[[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]]
[[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]]
-
*Venus
+
*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
[[Team:Chiba/protocol/transformation|Transformation]]
[[Team:Chiba/protocol/transformation|Transformation]]
-
*Venus
+
*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
*PUC19
*PUC19
*pGFPUV
*pGFPUV
Line 238: Line 239:
*Membren Venus YFP
*Membren Venus YFP
[[Team:Chiba/protocol/PCR|PCR]]
[[Team:Chiba/protocol/PCR|PCR]]
-
*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001]
+
*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001](1)
<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">No.</td>
-
<td></td>
+
<td>1</td>
</tr>
</tr>
<tr>
<tr>
-
<td>DNA tamplate</td>
+
<td>DNA tamplate(&mu;L)</td>
<td>1</td>
<td>1</td>
</tr>
</tr>
<tr>
<tr>
-
<td>FW primer(Venus)</td>
+
<td>FW primer(Venus)(&mu;L)</td>
<td>5</td>
<td>5</td>
</tr>
</tr>
<tr>
<tr>
-
<td>VR primer</td>
+
<td>VR primer(&mu;L)</td>
<td>5</td>
<td>5</td>
</tr>
</tr>
<tr>
<tr>
-
<td>dNTPmix</td>
+
<td>dNTPmix(&mu;L)</td>
<td>10</td>
<td>10</td>
</tr>
</tr>
<tr>
<tr>
-
<td>Thermo pol buffer</td>
+
<td>Thermo pol buffer(&mu;L)</td>
<td>10</td>
<td>10</td>
</tr>
</tr>
<tr>
<tr>
-
<td>DNA pol(VENT)</td>
+
<td>DNA pol(VENT)(&mu;L)</td>
<td>1</td>
<td>1</td>
</tr>
</tr>
<tr>
<tr>
-
<td>dH<sub>2</sub>O</td>
+
<td>dH<sub>2</sub>O(&mu;L)</td>
<td>68</td>
<td>68</td>
</tr>
</tr>
<tr>
<tr>
-
<td>TOTAL</td>
+
<td>TOTAL(&mu;L)</td>
<td>100</td>
<td>100</td>
</tr>
</tr>
</table>
</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.

[http://partsregistry.org/Part:BBa_J22136 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:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007)
  2. Double Digesiton:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
  3. Single Digestion:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
  4. [http://partsregistry.org/Part:BBa_R0010 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:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007) -> OK
  2. Double Digesiton:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007) -> ?(low)
  3. Single Digestion:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007) -> ?(low)
  4. [http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007) -> OK(low)


--->Digestion (Double Digestion)

  1. [http://partsregistry.org/Part:BBa_C0178 BBa_C0178]
  2. [http://partsregistry.org/Part:BBa_C0170 BBa_C0170]
  3. [http://partsregistry.org/Part:BBa_J04500 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

  • [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)
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

  • [http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
  • [http://partsregistry.org/Part:BBa_F2620 BBa_F2620]


Transformation

  • [http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
  • PUC19
  • pGFPUV
  • pGEX Venus YFP
  • Membren Venus YFP

PCR

  • [http://partsregistry.org/Part:BBa_J63001 BBa_J63001](1)
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