Team:Chiba/Calendar-Home/28 August 2008

From 2008.igem.org

(Difference between revisions)
(New page: '''OUTPUT TEAM''' <br>TIME RESPONSE 測定 <br>pGFP <br>sfGFP <br>LacZ <br>Venus(PGEX) <br>contol)
Line 1: Line 1:
-
'''OUTPUT TEAM'''
+
>[[Team:Chiba|Home]] | [[Team:Chiba/Notebook|Notebook]]
-
<br>TIME RESPONSE 測定
+
 
-
<br>pGFP
+
[[Team:Chiba/Calendar-Home/18 August 2008|18 August 2008 <]]|[[Team:Chiba/Calendar-Home/20 August 2008|> 20 August 2008]]
-
<br>sfGFP
+
 
-
<br>LacZ
+
==Laboratory work==
-
<br>Venus(PGEX)
+
===Team:Input===
-
<br>contol
+
'''[[Team:Chiba/protocol/transformation|Transformation]]'''
 +
 
 +
[http://partsregistry.org/Part:BBa_I7106 BBa_I7106](2007)
 +
生えてきたコロニーを10ml培養して'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''(60mlで溶出):
 +
 
 +
-->'''[[Team:Chiba/protocol/digestion|Digestion]]'''
 +
<br>混ぜ表
 +
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<td width="257"></td>
 +
<td>double</td>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
 +
<td>2</td>
 +
</tr>
 +
<tr>
 +
<td>10×BSA</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>10×NE</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>EcoRI</td>
 +
<td>0.5</td>
 +
</tr>
 +
<tr>
 +
<td>PstI</td>
 +
<td>0.5</td>
 +
</tr>
 +
<tr>
 +
<td>DNA</td>
 +
<td>5</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>10</td>
 +
</tr>
 +
</table>
 +
'''[[Team:Chiba/protocol/digestion|Digestion]]''' 37℃、30分
 +
<BR>プラスミドの濃度は33.6ng/μl
 +
 
 +
 
 +
 
 +
'''[[Team:Chiba/protocol/PCR|PCR]]'''
 +
<BR>RBS＋CI
 +
<BR>混ぜ表
 +
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">
 +
</tr>
 +
<tr>
 +
<td>DNA</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>FW primer</td>
 +
<td>2.5</td>
 +
</tr>
 +
<tr>
 +
<td>VR primer</td>
 +
<td>2.5</td>
 +
</tr>
 +
<tr>
 +
<td>dNTPmix</td>
 +
<td>5</td>
 +
</tr>
 +
<tr>
 +
<td>thermo pol buffer</td>
 +
<td>5</td>
 +
</tr>
 +
<tr>
 +
<td>DNA pol(VENT)</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
 +
<td>34</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>50</td>
 +
</tr>
 +
</table>
 +
 
 +
 
 +
'''[[Team:Chiba/protocol/gelcheck|ゲルチェック]]'''
 +
 
 +
 
 +
 
 +
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<td width="100">PCR産物</td>
 +
<td>1</td>
 +
<tr>
 +
<td>Dye</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>dH<SUB>2</SUB>O</td>
 +
<td>4</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>6</td>
 +
</tr>
 +
</table>
 +
 
 +
-->濃度は33.6ng/μg
 +
 
 +
 
 +
ベクターの'''[[Team:Chiba/protocol/PCR|PCR]]'''
 +
<br>[http://partsregistry.org/Part:BBa_I7106 BBa_I7106]
 +
<br>混ぜ表
 +
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">
 +
</tr>
 +
<tr>
 +
<td>DNA</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>FW primer</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>
 +
 
 +
 
 +
'''[[Team:Chiba/protocol/gelcheck|ゲルチェック]]'''
 +
 
 +
             
 +
 
 +
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<td width="100">PCR産物</td>
 +
<td>1</td>
 +
<tr>
 +
<td>Dye</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
 +
<td>4</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>6</td>
 +
</tr>
 +
</table>
 +
-->バンド出ず。
 +
 
 +
 
 +
===Team:Communication===
 +
:(27/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]
 +
::#insert:C0170 + vector:J04500
 +
::#insert:C0178 + vector:J04500
 +
 
 +
 
 +
:--->'''[[Team:Chiba/protocol/digestion|Digestion Test]]'''
 +
::#insert:C0170 + vector:J04500
 +
::#insert:C0178 + vector:J04500
 +
 
 +
<table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample No.</td>
 +
<td>3</td><td>4</td><td>5</td><td>6</td>
 +
</tr>
 +
<tr>
 +
<td>insert:C0170 + vector:J04500 </td>
 +
<td>1</td><td>-</td><td>1</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>insert:C0178 + vector:J04500 </td>
 +
<td>-</td><td>1</td><td>-</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>EcoRⅠ</td>
 +
<td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
 +
</tr>
 +
<tr>
 +
<td>SpeⅠ</td>
 +
<td>-</td><td>-</td><td>0.1</td><td>0.1</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 3</td>
 +
<td>1</td><td>1</td><td>1</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>BSA</td>
 +
<td>-</td><td>-</td><td>1</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
 +
<td>7.8</td><td>7.8</td><td>6.8</td><td>6.8</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>10</td><td>10</td><td>10</td><td>10</td>
 +
</tr>
 +
</table>
 +
 
 +
 
 +
 
 +
:--->(29/8)'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
 +
{|align="justify"
 +
|[[Image:Chiba-0828.JPG]]
 +
|
 +
:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample No.</td>
 +
<td>1,2</td><td>3~6</td>
 +
</tr>
 +
<tr>
 +
<td>Sample DNA</td>
 +
<td>1</td><td>10</td>
 +
</tr>
 +
<tr>
 +
<td>Loading Dye</td>
 +
<td>1</td><td>2</td>
 +
</tr>
 +
<tr>
 +
<td>H2O</td>
 +
<td>4</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>6</td><td>12</td>
 +
</tr>
 +
</table>
 +
:From left;
 +
::#　-> OK
 +
::#　-> OK
 +
::#　-> OK
 +
::#　-> OK
 +
::#　-> OK
 +
::#　-> OK
 +
|}
 +
 
 +
 
 +
===Team:Output===
 +
[[Team:Chiba/jk/γ/Trl|Time Responce(liquid)]]

Revision as of 07:11, 24 October 2008

>Home | Notebook

18 August 2008 <|> 20 August 2008

Contents

Laboratory work

Team:Input

Transformation

[http://partsregistry.org/Part:BBa_I7106 BBa_I7106](2007) 生えてきたコロニーを10ml培養してMini prep(60mlで溶出):

-->Digestion
混ぜ表

double
dH2O 2
10×BSA 1
10×NE 1
EcoRI 0.5
PstI 0.5
DNA 5
TOTAL 10

Digestion 37℃、30分
プラスミドの濃度は33.6ng/μl


PCR
RBS＋CI
混ぜ表

DNA 1
FW primer 2.5
VR primer 2.5
dNTPmix 5
thermo pol buffer 5
DNA pol(VENT) 1
dH2O 34
TOTAL 50


ゲルチェック


PCR産物 1
Dye 1
dH2O 4
TOTAL 6

-->濃度は33.6ng/μg


ベクターのPCR
[http://partsregistry.org/Part:BBa_I7106 BBa_I7106]
混ぜ表

DNA 1
FW primer 5
VR primer 5
dNTPmix 10
thermo pol buffer 10
DNA pol(VENT) 1
dH2O 68
TOTAL 100


ゲルチェック


PCR産物 1
Dye 1
dH2O 4
TOTAL 6

-->バンド出ず。


Team:Communication

(27/8)--->Mini prep
  1. insert:C0170 + vector:J04500
  2. insert:C0178 + vector:J04500


--->Digestion Test
  1. insert:C0170 + vector:J04500
  2. insert:C0178 + vector:J04500
Sample No. 3456
insert:C0170 + vector:J04500 1-1-
insert:C0178 + vector:J04500 -1-1
EcoRⅠ 0.10.10.10.1
SpeⅠ --0.10.1
Buffer 3 1111
BSA --11
dH2O 7.87.86.86.8
TOTAL 10101010


--->(29/8)Gel Check
Chiba-0828.JPG
Sample No. 1,23~6
Sample DNA 110
Loading Dye 12
H2O 4-
TOTAL 612
From left;
  1. 　-> OK
  2. 　-> OK
  3. 　-> OK
  4. 　-> OK
  5. 　-> OK
  6. 　-> OK


Team:Output

Time Responce(liquid)

Retrieved from "http://2008.igem.org/Team:Chiba/Calendar-Home/28_August_2008"