Team:Chiba/jk/γ/week 2

From 2008.igem.org

(Difference between revisions)
(30,August,2008)
(26,August,2008)
 
(5 intermediate revisions not shown)
Line 1: Line 1:
[[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 2 ==
== WEEK 2 ==
== 24,August 2008 ==
== 24,August 2008 ==
 +
[[Team:Chiba/protocol/transformation|Transformation]]
 +
*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001](yeast optimized YFP)-->colony
 +
 +
[[Team:Chiba/protocol/digestion|Digestion]]
 +
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](1)①
 +
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](2)②
 +
*Menbren Venus YFP③
-
<br>TRANSFORMATION
 
-
<br>BBa_J63001(yeast optimized YFP)-->colony
 
-
<br>Digestion
 
-
<br>BBa_F2620(1)①
 
-
<br>BBa_F2620(2)②
 
-
<br>Menbren Venus YFP③
 
<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>③</td>
<td>①</td><td>②</td><td>③</td>
</tr>
</tr>
Line 26: Line 28:
</tr>
</tr>
<tr>
<tr>
-
<td>dH2O</td>
+
<td>dH<sub>2</sub>O</td>
<td>6</td><td>6</td><td>6</td>
<td>6</td><td>6</td><td>6</td>
</tr>
</tr>
Line 38: Line 40:
</tr>
</tr>
</table>
</table>
-
<br>->37℃ 30min
+
 
 +
 
 +
-->37℃ 30min
 +
 
 +
 
<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(afetr digestion)</td><td>②(after digestion)</td><td>③(afetr digestion)</td>
<td>①(afetr digestion)</td><td>②(after digestion)</td><td>③(afetr digestion)</td>
</tr>
</tr>
Line 57: Line 63:
</tr>
</tr>
</table>
</table>
 +
 +
<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>③</td>
<td>①</td><td>②</td><td>③</td>
</tr>
</tr>
Line 71: Line 79:
</tr>
</tr>
<tr>
<tr>
-
<td>dH2O</td>
+
<td>dH<sub>2</sub>O</td>
<td>3</td><td>3</td><td>3</td>
<td>3</td><td>3</td><td>3</td>
</tr>
</tr>
Line 79: Line 87:
</tr>
</tr>
</table>
</table>
-
<br>->135V 30min
+
 
 +
-->135V 30min
== 25,August,2008 ==
== 25,August,2008 ==
-
<br>PCR
+
[[Team:Chiba/protocol/PCR|PCR]]
-
<br>BBa_E2030①
+
*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]①
-
<br>BBa_J33202②
+
*[http://partsregistry.org/Part:BBa_J33202 BBa_J33202]②
-
<br>BL21③
+
*BL21③
-
<br>BBa_J52008④
+
*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]④
-
<br>BBa_I712019⑤
+
*[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">Sample No.</td>
<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
</tr>
</tr>
Line 120: Line 130:
</tr>
</tr>
<tr>
<tr>
-
<td>dH2O</td>
+
<td>dH<sub>2</sub>O</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>
Line 128: Line 138:
</tr>
</tr>
</table>
</table>
-
<br>->Gel  
+
 
 +
-->[[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</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
</tr>
</tr>
Line 143: Line 156:
</tr>
</tr>
<tr>
<tr>
-
<td>dH2O</td>
+
<td>dH<sub>2</sub>O</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>
Line 152: Line 165:
</table>
</table>
-
<br>MINI prep
+
[[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]]
-
<br>Venus
+
*Venus
-
<br>BBa_F2620
+
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
-
<br>TRANSFORMATRION
+
 
-
<br>Venus
+
 
-
<br>PUC19
+
[[Team:Chiba/protocol/transformation|Transformation]]
-
<br>pGFPUV
+
*Venus
-
<br>pGEX Venus YFP
+
*PUC19
-
<br>Membren Venus YFP
+
*pGFPUV
-
<br>PCR
+
*pGEX Venus YFP
-
<br>BBa_J63001①
+
*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">
<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 193: Line 208:
</tr>
</tr>
<tr>
<tr>
-
<td>dH2O</td>
+
<td>dH<sub>2</sub>O</td>
<td>68</td>
<td>68</td>
</tr>
</tr>
Line 204: Line 219:
== 26,August,2008 ==
== 26,August,2008 ==
-
<br>Gel check
+
[[Team:Chiba/protocol/gelcheck|Gel Check]]
-
<br>BBa_F2620
+
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
-
<br>BBa_F2620
+
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
-
<br>BBa_J63001
+
*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001]
<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>③</td><td>④</td><td>⑤</td>
<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
</tr>
</tr>
Line 222: Line 237:
</tr>
</tr>
<tr>
<tr>
-
<td>dH2O</td>
+
<td>dH<sub>2</sub>O</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>
Line 231: Line 246:
</table>
</table>
-
<br>Digestion
+
[[Team:Chiba/protocol/digestion|Digestion]]
-
<br>vector:BBa_F2620
+
*vector:[http://partsregistry.org/Part:BBa_F2620 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">Sample No.</td>
-
<td>BBa_F2620</td>
+
<td>[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]</td>
</tr>
</tr>
<tr>
<tr>
Line 258: Line 274:
<td>13</td>
<td>13</td>
</tr>
</tr>
 +
<tr>
<td>TOTAL</td>
<td>TOTAL</td>
<td>130</td>
<td>130</td>
</tr>
</tr>
</table>
</table>
-
<br>insert:BBa_J52008,BBa_E2030
+
 
 +
*insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008],[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
 +
 
<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>J52008</td><td>E2030</td>
+
<td>[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]</td><td>[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]</td>
</tr>
</tr>
<tr>
<tr>
Line 292: Line 311:
<td>4</td><td>4</td>
<td>4</td><td>4</td>
</tr>
</tr>
 +
<tr>
<td>TOTAL</td>
<td>TOTAL</td>
<td>40</td><td>40</td>
<td>40</td><td>40</td>
</tr>
</tr>
</table>
</table>
-
<br>->37℃ 3hour
+
<br>-->37℃ 3hour
-
<br>->SAP
+
<br>-->SAP
-
<br>->37℃ 30min
+
<br>-->37℃ 30min
-
<br>->65℃ 15min
+
<br>-->65℃ 15min
-
<br>->Zymo Clean
+
<br>-->[[Team:Chiba/protocol/DNA_Purification/zymocleam|Zymo Clean]]
-
<br>Ligation
+
 
-
<br>vector:BBa_F2620 insert:BBa_J52008①
+
[[Team:Chiba/protocol/ligation/ligation|Ligation]]
-
<br>vector:BBa_F2620 insert:BBa_E2030②
+
 
-
<br>Negative control:BBa_F2620③
+
*vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]①
 +
*vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]②
 +
*Negative control:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]③
 +
 
<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>③</td>
<td>①</td><td>②</td><td>③</td>
</tr>
</tr>
Line 327: Line 350:
</tr>
</tr>
<tr>
<tr>
-
<td>dH2O</td>
+
<td>dH<sub>2</sub>O</td>
<td>0</td><td>2.5</td><td>10</td>
<td>0</td><td>2.5</td><td>10</td>
</tr>
</tr>
Line 335: Line 358:
</tr>
</tr>
</table>
</table>
-
<br>->RT 2hour
 
-
<br>->TRANSFORMATION XL10G
 
-
<br>->37℃ over night
 
-
<br>->colony PCR
 
-
<br>->MINI PREP
 
 +
-->RT 2hour
 +
 +
-->[[Team:Chiba/protocol/transformation|Transformation]](XL10G)
 +
 +
-->37℃ over night
 +
 +
-->colony PCR
 +
 +
-->[[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]]
== 28,August,2008 ==
== 28,August,2008 ==
-
<br>[[Team:Chiba/jk/γ/Trl|Time Responce(liquid)]]
+
 
 +
[[Team:Chiba/jk/γ/Trl|Time Responce(liquid)]]
Line 350: Line 378:
== 29,August,2008 ==
== 29,August,2008 ==
-
<br>[[Team:Chiba/jk/γ/trs|TIME RESPONCE(solid)]]
+
 
 +
[[Team:Chiba/jk/γ/trs|TIME RESPONCE(solid)]]
== 30,August,2008 ==
== 30,August,2008 ==
-
<br>TRANSFORMATION
+
[[Team:Chiba/protocol/transformation|Transformation]]
-
<br>mcherry
+
 
-
<br>PCR
+
*mcherry
-
<br>BBa_J52008(rLuc)
+
 
 +
[[Team:Chiba/protocol/PCR|PCR]]
 +
 
 +
*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](rLuc)
 +
 
<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>BBa_J52008</td>
+
<td>[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]</td>
</tr>
</tr>
<tr>
<tr>
Line 389: Line 422:
</tr>
</tr>
<tr>
<tr>
-
<td>dH2O</td>
+
<td>dH<sub>2</sub>O</td>
<td>34</td>
<td>34</td>
</tr>
</tr>
 +
<tr>
<td>TOTAL</td>
<td>TOTAL</td>
<td>50</td>
<td>50</td>
Line 401: Line 435:
[[Team:Chiba/jk/γ/week 1|>Last week]]
[[Team:Chiba/jk/γ/week 1|>Last week]]
-
<br>[[Team:Chiba/jk/γ/week 3|>Next week]]
+
 
 +
[[Team:Chiba/jk/γ/week 3|>Next week]]

Latest revision as of 05:50, 24 October 2008

>実験ログ

>出力班

Contents

WEEK 2

24,August 2008

Transformation

  • [http://partsregistry.org/Part:BBa_J63001 BBa_J63001](yeast optimized YFP)-->colony

Digestion

  • [http://partsregistry.org/Part:BBa_F2620 BBa_F2620](1)①
  • [http://partsregistry.org/Part:BBa_F2620 BBa_F2620](2)②
  • Menbren Venus YFP③
Sample No.
DNA tamplate 333
Buffer3 111
dH2O 666
EcoRⅠ 0.50.50.5
TOTAL 101010


-->37℃ 30min


Sample No. ①(afetr digestion)②(after digestion)③(afetr digestion)
DNA tamplate 101010
loading Dye 222
TOTAL 121212


Sample No.
DNA tamplate 222
loading Dye 111
dH2O 333
TOTAL 666

-->135V 30min


25,August,2008

PCR

  • [http://partsregistry.org/Part:BBa_E2030 BBa_E2030]①
  • [http://partsregistry.org/Part:BBa_J33202 BBa_J33202]②
  • BL21③
  • [http://partsregistry.org/Part:BBa_J52008 BBa_J52008]④
  • [http://partsregistry.org/Part:BBa_I712019 BBa_I712019]⑤
>
Sample No.
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.
DNA tamplate 11111
loading Dye 11111
dH2O 44444
TOTAL 66666

Mini prep

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


Transformation

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

PCR

  • [http://partsregistry.org/Part:BBa_J63001 BBa_J63001]①
Sample No.
DNA tamplate 1
FW primer(Venus) 5
VR primer 5
dNTPmix 10
Thermo pol buffer 10
DNA pol(VENT) 1
dH2O 68
TOTAL 100

26,August,2008

Gel Check

  • [http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
  • [http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
  • [http://partsregistry.org/Part:BBa_J63001 BBa_J63001]
>
Sample No.
DNA tamplate 11111
Loading Dye 11111
dH2O 44444
TOTAL 66666

Digestion

  • vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
Sample No. [http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
DNA tamplate 100
SpeⅠ 2
PstⅠ 2
BSA(x10) 13
Buffer2 13
TOTAL 130
  • insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008],[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
Sample No. [http://partsregistry.org/Part:BBa_J52008 BBa_J52008][http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
DNA tamplate 3030
XbaⅠ 11
PstⅠ 11
DpnⅠ 11
Buffer3 44
BSA(x10) 44
TOTAL 4040


-->37℃ 3hour
-->SAP
-->37℃ 30min
-->65℃ 15min
-->Zymo Clean

Ligation

  • vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]①
  • vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]②
  • Negative control:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]③
Sample No.
vector 555
inser 107.50
Ligase Buffer 444
Ligase 111
dH2O 02.510
TOTAL 202020

-->RT 2hour

-->Transformation(XL10G)

-->37℃ over night

-->colony PCR

-->Mini prep

28,August,2008

Time Responce(liquid)


29,August,2008

TIME RESPONCE(solid)


30,August,2008

Transformation

  • mcherry

PCR

  • [http://partsregistry.org/Part:BBa_J52008 BBa_J52008](rLuc)
Sample No. [http://partsregistry.org/Part:BBa_J52008 BBa_J52008]
DNA tamplate 1
rLuc_fwd 2.5
Rev primer 2.5
Thermo pol Buffer 5
dNTPmix 5
Vent pol 0.5
dH2O 34
TOTAL 50



>Last week

>Next week


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