Team:Chiba/Calendar-Home/28 August 2008

From 2008.igem.org

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(New page: '''OUTPUT TEAM''' <br>TIME RESPONSE 測定 <br>pGFP <br>sfGFP <br>LacZ <br>Venus(PGEX) <br>contol)
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'''OUTPUT TEAM'''
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>[[Team:Chiba|Home]] | [[Team:Chiba/Notebook|Notebook]]
-
<br>TIME RESPONSE 測定
+
 
-
<br>pGFP
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[[Team:Chiba/Calendar-Home/27 August 2008|27 August 2008 <]]|[[Team:Chiba/Calendar-Home/29 August 2008|> 29 August 2008]]
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<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)
 +
'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
 +
 
 +
-->'''[[Team:Chiba/protocol/digestion|Digestion]]'''
 +
<br>
 +
<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&mu;l </td>
 +
</tr>
 +
</table>
 +
'''[[Team:Chiba/protocol/digestion|Digestion]]''' 37&deg;C、30min
 +
<BR>-->Concentration 33.6ng/&mu;l
 +
 
 +
 
 +
 
 +
'''[[Team:Chiba/protocol/PCR|PCR]]'''
 +
<BR>RBS＋CI
 +
<BR>
 +
<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&mu;l </td>
 +
</tr>
 +
</table>
 +
 
 +
 
 +
'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
 +
 
 +
 
 +
 
 +
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<td width="100">sampleDNA</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>6μl </td>
 +
</tr>
 +
</table>
 +
 
 +
-->Concentration:33.6ng/μl
 +
 
 +
 
 +
Vector'''[[Team:Chiba/protocol/PCR|PCR]]'''
 +
<br>[http://partsregistry.org/Part:BBa_I7106 BBa_I7106]
 +
<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>100μl </td>
 +
</tr>
 +
</table>
 +
 
 +
 
 +
'''[[Team:Chiba/protocol/gelcheck|Gel check]]'''
 +
 
 +
             
 +
 
 +
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<td width="100">sampleDNA</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>6μl </td>
 +
</tr>
 +
</table>
 +
-->none
 +
 
 +
===Team:Communication===
 +
:(27/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]
 +
::#insert:[http://partsregistry.org/Part:BBa_C0170 BBa_C0170] + vector:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]
 +
::#insert:[http://partsregistry.org/Part:BBa_C0178 BBa_C0178] + vector:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]
 +
 
 +
 
 +
:--->'''[[Team:Chiba/protocol/digestion|Digestion Test]]'''
 +
::#insert:[http://partsregistry.org/Part:BBa_C0170 BBa_C0170] + vector:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]
 +
::#insert:[http://partsregistry.org/Part:BBa_C0178 BBa_C0178] + vector:[http://partsregistry.org/Part:BBa_J04500 BBa_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>10μl</td><td>10μl</td><td>10μl</td><td>10μl</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>H<sub>2</sub>O</td>
 +
<td>4</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>6μl</td><td>12μl</td>
 +
</tr>
 +
</table>
 +
:From left;
 +
::#　-> OK
 +
::#　-> OK
 +
::#　-> OK
 +
::#　-> OK
 +
::#　-> OK
 +
::#　-> OK
 +
|}
 +
 
 +
===Team:Output===
 +
[[Team:Chiba/jk/γ/Trl|Time Responce(liquid)]]

Latest revision as of 03:14, 30 October 2008

>Home | Notebook

27 August 2008 <|> 29 August 2008

Contents

Laboratory work

Team:Input

Transformation

[http://partsregistry.org/Part:BBa_I7106 BBa_I7106](2007) Mini prep

-->Digestion

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

Digestion 37°C、30min
-->Concentration 33.6ng/μl


PCR
RBS＋CI

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


Gel Check


sampleDNA 1
Dye 1
dH2O 4
TOTAL 6μl

-->Concentration:33.6ng/μl


VectorPCR
[http://partsregistry.org/Part:BBa_I7106 BBa_I7106]

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


Gel check


sampleDNA 1
Dye 1
dH2O 4
TOTAL 6μl

-->none

Team:Communication

(27/8)--->Mini prep
  1. insert:[http://partsregistry.org/Part:BBa_C0170 BBa_C0170] + vector:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]
  2. insert:[http://partsregistry.org/Part:BBa_C0178 BBa_C0178] + vector:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]


--->Digestion Test
  1. insert:[http://partsregistry.org/Part:BBa_C0170 BBa_C0170] + vector:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]
  2. insert:[http://partsregistry.org/Part:BBa_C0178 BBa_C0178] + vector:[http://partsregistry.org/Part:BBa_J04500 BBa_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 10μl10μl10μl10μl


--->(29/8)Gel Check
Chiba-0828.JPG
Sample No. 1,23~6
Sample DNA 110
Loading Dye 12
H2O 4-
TOTAL 6μl12μl
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"