Team:Evry/Notebook/w7
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<h2>Tuesday, 24th June</h2> | <h2>Tuesday, 24th June</h2> | ||
+ | <h3>Plasmid Purification:</h3> | ||
+ | On the <strong>pCS2+ RFP</strong> clones incubated the day before: | ||
+ | |||
+ | <TABLE BORDER="1"> | ||
+ | <CAPTION> DNA Concentration </CAPTION> | ||
+ | <TR> | ||
+ | <TH> <center> Tube </center></TH> | ||
+ | <TH> <center> Concentration (ng.uL-1) </center></TH> | ||
+ | </TR> | ||
+ | <TR> | ||
+ | <TH> 1 </TH> | ||
+ | <TD> <center>144,2</center></TD> | ||
+ | </TR> | ||
+ | <TR> | ||
+ | <TH> 2 </TH> | ||
+ | <TD> <center>161,9</center></TD> | ||
+ | </TR> | ||
+ | <TR> | ||
+ | <TH> 3 </TH> | ||
+ | <TD> <center>184,9</center></TD> | ||
+ | </TR> | ||
+ | <TR> | ||
+ | <TH> 4 </TH> | ||
+ | <TD> <center>120,7</center></TD> | ||
+ | </TR> | ||
+ | </TABLE> | ||
+ | |||
+ | <h3>Digestion:</h3> | ||
+ | |||
+ | <TABLE BORDER="1"> | ||
+ | <CAPTION> Plasmid digestion </CAPTION> | ||
+ | <TR> | ||
+ | <TH> <center> Tube </center></TH> | ||
+ | <TH> <center> V DNA (uL) </center></TH> | ||
+ | <TH> <center> V SpeI (uL) </center></TH> | ||
+ | <TH> <center> V EcoRI (uL) </center></TH> | ||
+ | <TH> <center> V buffer (uL) </center></TH> | ||
+ | <TH> <center> V H2O (uL) </center></TH> | ||
+ | </TR> | ||
+ | <TR> | ||
+ | <TH> 1 </TH> | ||
+ | <TD> <center>7</center></TD> | ||
+ | <TD> <center>1</center></TD> | ||
+ | <TD> <center>1</center></TD> | ||
+ | <TD> <center>2</center></TD> | ||
+ | <TD> <center>9</center></TD> | ||
+ | </TR> | ||
+ | <TR> | ||
+ | <TH> 2 </TH> | ||
+ | <TD> <center>6,2</center></TD> | ||
+ | <TD> <center>1</center></TD> | ||
+ | <TD> <center>1</center></TD> | ||
+ | <TD> <center>2</center></TD> | ||
+ | <TD> <center>9,8</center></TD> | ||
+ | </TR> | ||
+ | <TR> | ||
+ | <TH> 3 </TH> | ||
+ | <TD> <center>5,4</center></TD> | ||
+ | <TD> <center>1</center></TD> | ||
+ | <TD> <center>1</center></TD> | ||
+ | <TD> <center>2</center></TD> | ||
+ | <TD> <center>10,6</center></TD> | ||
+ | </TR> | ||
+ | <TR> | ||
+ | <TH> 4 </TH> | ||
+ | <TD> <center>8,3</center></TD> | ||
+ | <TD> <center>1</center></TD> | ||
+ | <TD> <center>1</center></TD> | ||
+ | <TD> <center>2</center></TD> | ||
+ | <TD> <center>7,7</center></TD> | ||
+ | </TR> | ||
+ | </TABLE> | ||
+ | |||
+ | <h3>Gel migration:</h3> | ||
+ | |||
+ | Gel 0,8% | ||
<h2>Thursday, 26th June</h2> | <h2>Thursday, 26th June</h2> |
Revision as of 21:09, 2 August 2012
Weeks
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 |
Week 7: 23rd July - 29th July
Monday, 23rd June
Cloning:
4 different clones pCS2+ RFP => incubation in LB medium overnight at 37 degree celsius.PCR:
PCR of pCS2+:Reactants | Volumes (µl) | ||||
GC Buffer | 10 | ||||
dNTPs | 1 | ||||
DNA | 1 | ||||
H2O | 32,5 | ||||
Primers (FW and RV) | 2,5 each | ||||
fusion polymerase | 0,5 |
Tuesday, 24th June
Plasmid Purification:
On the pCS2+ RFP clones incubated the day before: |
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Digestion:
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