Team:Evry/Notebook/July/6

From 2012.igem.org

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<center><h1>Promoters & Reporters workgroup</h1></center>
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<FONT SIZE=3>'''Digestion'''</FONT> <br/>
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<br/>
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Miniprep 28/06/2012<br/>
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<br/>
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masse DNA = 2000 ng
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Volume final = 30 uL
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<TABLE BORDER="1">
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  <TR>
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    <TH> <center> DNA </center></TH>
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    <TH> <center>Concentration (ng.uL-1) </center></TH>
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    <TH> <center> V DNA (uL)</center></TH>
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    <TH> <center> V SpeI (uL)</center></TH>
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    <TH> <center> V EcoRI (uL)</center></TH>
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    <TH> <center> V XBA I (uL)</center></TH>
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    <TH> <center> V Buffer Orange (uL)</center></TH>
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    <TH> <center> V Buffer sup (uL)</center></TH>
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    <TH> <center> V water (uL)</center></TH>
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  </TR>
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  <TR>
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    <TH> <center>pCS2 +</center></TH>
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    <TD> <center> 362,7 </center> </TD>
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    <TD> <center> 5,51 </center> </TD>
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    <TD> <center> 0 </center> </TD>
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    <TD> <center> 1 </center> </TD>
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    <TD> <center> 1 </center> </TD>
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    <TD> <center> 3 </center> </TD>
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    <TD> <center> 0 </center> </TD>
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    <TD> <center> 19,49 </center> </TD>
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  </TR>
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  <TR>
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    <TH> CFP </TH>
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    <TD> <center> 240,1</center> </TD>
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    <TD> <center> 8,33 </center> </TD>
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    <TD> <center> 1 </center> </TD>
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    <TD> <center> 1 </center> </TD>
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    <TD> <center> 0 </center> </TD>
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    <TD> <center> 0 </center> </TD>
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    <TD> <center> 3 </center> </TD>
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    <TD> <center> 16,67 </center> </TD>
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  </TR>
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  <TR>
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    <TH> GFP </TH>
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    <TD> <center> 209,1</center> </TD>
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    <TD> <center> 9,56 </center> </TD>
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    <TD> <center> 1 </center> </TD>
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    <TD> <center> 1 </center> </TD>
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    <TD> <center> 0 </center> </TD>
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    <TD> <center> 0 </center> </TD>
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    <TD> <center> 3 </center> </TD>
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    <TD> <center> 15,44 </center> </TD>
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  </TR>
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  <TR>
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    <TH> RFP </TH>
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    <TD> <center> 142,8</center> </TD>
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    <TD> <center> 14,01 </center> </TD>
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    <TD> <center> 1 </center> </TD>
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    <TD> <center> 1 </center> </TD>
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    <TD> <center> 0 </center> </TD>
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    <TD> <center> 0 </center> </TD>
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    <TD> <center> 3 </center> </TD>
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    <TD> <center> 10,99 </center> </TD>
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  </TR>
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  <TR>
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    <TH> YFP </TH>
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    <TD> <center> 196,0</center> </TD>
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    <TD> <center> 10,20 </center> </TD>
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    <TD> <center> 1 </center> </TD>
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    <TD> <center> 1 </center> </TD>
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    <TD> <center> 0 </center> </TD>
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    <TD> <center> 0 </center> </TD>
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    <TD> <center> 3 </center> </TD>
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    <TD> <center> 14,80 </center> </TD>
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  </TR>
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</TABLE>
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<br/>
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Incubation 3h at 37 degrees Celsius<br/>
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<br/>
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<FONT SIZE=3>'''Gel Migration'''</FONT> <br/>
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<br/>
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Gel at 0,08% <br/>
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V tae = 30 mL<br/>
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m aga = 0,24 g <br/>
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V bet = 3 uL<br/>
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<br/>
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No Band for RFP (again)<br/>
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<br/>
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<FONT SIZE=3>'''Gel Extraction'''</FONT> <br/>
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<br/>
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see protocol on 05/07/2012
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<TABLE BORDER="1">
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  <TR>
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    <TH> <center> Type </center></TH>
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    <TH> <center>Gel weight (g) </center></TH>
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    <TH> <center>V QG (uL) </center></TH>
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    <TH> <center>V iso (uL) </center></TH>
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  </TR>
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  <TR>
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    <TH> pCS2 </TH>
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    <TD> <center>0,37</center></TD>
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    <TD> <center>1 110</center></TD>
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    <TD> <center>370</center></TD>
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  </TR>
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  <TR>
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    <TH> YFP </TH>
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    <TD> <center>0,17</center></TD>
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    <TD> <center>510</center></TD>
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    <TD> <center>170</center></TD>
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  </TR>
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  <TR>
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    <TH> GFP </TH>
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    <TD> <center>0,23</center></TD>
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    <TD> <center>690</center></TD>
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    <TD> <center>230</center></TD>
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  </TR>
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  <TR>
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    <TH> CFP </TH>
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    <TD> <center>0,19</center></TD>
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    <TD> <center>570</center></TD>
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    <TD> <center>190</center></TD>
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  </TR>
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</TABLE>
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<br/>
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<TABLE BORDER="1">
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  <CAPTION> DNA Concentration </CAPTION>
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  <TR>
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    <TH> <center> Type </center></TH>
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    <TH> <center> Concentration (ng.uL-1) </center></TH>
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  </TR>
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  <TR>
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    <TH> pCS2 </TH>
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    <TD> <center>16,7</center></TD>
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  </TR>
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  <TR>
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    <TH> GFP </TH>
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    <TD> <center>8,5</center></TD>
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  </TR>
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  <TR>
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    <TH> CFP </TH>
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    <TD> <center>9,1</center></TD>
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  </TR>
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  <TR>
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    <TH> YFP </TH>
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    <TD> <center>5,6</center></TD>
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  </TR>
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</TABLE>
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<br/>
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<html>
<center>
<center>
<a href="https://2012.igem.org/Team:Evry/Notebook/July/5"><img  src="https://static.igem.org/mediawiki/2012/e/ed/Previous_day_arrow.png"></a>
<a href="https://2012.igem.org/Team:Evry/Notebook/July/5"><img  src="https://static.igem.org/mediawiki/2012/e/ed/Previous_day_arrow.png"></a>

Revision as of 16:10, 12 July 2012

Promoters & Reporters workgroup

Digestion

Miniprep 28/06/2012

masse DNA = 2000 ng Volume final = 30 uL

DNA
Concentration (ng.uL-1)
V DNA (uL)
V SpeI (uL)
V EcoRI (uL)
V XBA I (uL)
V Buffer Orange (uL)
V Buffer sup (uL)
V water (uL)
pCS2 +
362,7
5,51
0
1
1
3
0
19,49
CFP
240,1
8,33
1
1
0
0
3
16,67
GFP
209,1
9,56
1
1
0
0
3
15,44
RFP
142,8
14,01
1
1
0
0
3
10,99
YFP
196,0
10,20
1
1
0
0
3
14,80


Incubation 3h at 37 degrees Celsius

Gel Migration

Gel at 0,08%
V tae = 30 mL
m aga = 0,24 g
V bet = 3 uL

No Band for RFP (again)

Gel Extraction

see protocol on 05/07/2012

Type
Gel weight (g)
V QG (uL)
V iso (uL)
pCS2
0,37
1 110
370
YFP
0,17
510
170
GFP
0,23
690
230
CFP
0,19
570
190


DNA Concentration
Type
Concentration (ng.uL-1)
pCS2
16,7
GFP
8,5
CFP
9,1
YFP
5,6


Retrieved from "http://2012.igem.org/Team:Evry/Notebook/July/6"