Difference between revisions of "Team:Stony Brook/Notebook/Cancer-W11"

(Created page with "{{Stony_Brook TryingFailing}} <html> <head> <title> Week 11: 9/5 - 9/11 </title> </head> <body> <div class="column full_size"> <div align="center"> <h1> Week 11: 9/5 - 9/11 <...")
 
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<div align="center">
 
<div align="center">
 
<h2 > 9/5 </h2>
 
<h2 > 9/5 </h2>
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</div>
 +
</div>
 +
 +
<div class="column full_size">
 +
<div align="center">
 +
<h4> Digest of CR-1 Plasmids </h4>
 +
<table>
 +
<tr>
 +
<th> Content </th>
 +
<th> Experimental ul </th>
 +
<th> Control ul </th>
 +
</tr>
 +
<tr>
 +
<td> H<sub>2</sub>O </td>
 +
<td> 15.5</td>
 +
<td> 16.5 </td>
 +
</tr>
 +
<tr>
 +
<td> Cutsmart </td>
 +
<td> 2.5 </td>
 +
<td> 2.5 </td>
 +
</tr>
 +
<tr>
 +
<td> XhoI </td>
 +
<td> 0.5 </td>
 +
<td> X </td>
 +
</tr>
 +
<tr>
 +
<td> EcoRI-HF </td>
 +
<td> 0.5</td>
 +
<td> X </td>
 +
</tr>
 +
<tr>
 +
<td> DNA </td>
 +
<td> 6 </td>
 +
<td> 6 </td>
 +
</tr>
 +
</table>
 +
 +
<br>
 +
<br>
 +
 +
<h4> PCR of Toggle 2 </h4>
 +
</div>
 +
</div>
 +
 +
<div class="column half_size">
 +
<div align="center">
 +
<h5> PCR </h5>
 +
<table>
 +
<tr>
 +
<th> Contents </th>
 +
<th> Trial 1 (ul) </th>
 +
<th> Trial 2 (ul) </th>
 +
</tr>
 +
<td> Q5 MasterMix </td>
 +
<td> 12.5 </td>
 +
<td> 12.5 </td>
 +
</tr>
 +
<tr>
 +
<td> H<sub>2</sub>O</td>
 +
<td> 6.5 </td>
 +
<td> 5.5 </td>
 +
</tr>
 +
<tr>
 +
<td> 10 uM F Primer </td>
 +
<td> 0.5 </td>
 +
<td> 1.0 </td>
 +
</tr>
 +
<tr>
 +
<td> 10 uM R Primer </td>
 +
<td> 0.5 </td>
 +
<td> 1.0 </td>
 +
</tr>
 +
<tr>
 +
<td> 100pg/ul Construct </td>
 +
<td> 5 </td>
 +
<td> 5 </td>
 +
</tr>
 +
</table>
 +
</div>
 +
</div>
 +
 +
<div class="column half_size">
 +
<div align="center">
 +
<h5> PCR Settings: 30 cycles </h5>
 +
<table>
 +
<tr>
 +
<th> Phase </th>
 +
<th> Temperature (°C) </th>
 +
<th> Time (sec)</th>
 +
</tr>
 +
<tr>
 +
<td> Initial Denaturation </td>
 +
<td> 98 </td>
 +
<td> 30 </td>
 +
</tr>
 +
<tr>
 +
<td> Denaturation </td>
 +
<td> 98 </td>
 +
<td> 10 </td>
 +
</tr>
 +
<tr>
 +
<td> Annealing </td>
 +
<td> 68 </td>
 +
<td> 20 </td>
 +
</tr>
 +
<tr>
 +
<td> Extension </td>
 +
<td> 72 </td>
 +
<td> 60 </td>
 +
</tr>
 +
<tr>
 +
<td> Final Extension </td>
 +
<td> 72 </td>
 +
<td> 120 </td>
 +
</tr>
 +
</table>
 +
</div>
 +
</div>
 +
 +
<div class="column full_size">
 +
<div align="center">
 +
 +
<h4> Ran Digests </h4>
 +
<br>
 +
 +
<h5> Digest of J6100 </h5>
 +
<table>
 +
<tr>
 +
<th> Content </th>
 +
<th> Experimental ul </th>
 +
</tr>
 +
<tr>
 +
<td> H<sub>2</sub>O </td>
 +
<td> 7.5 </td>
 +
</tr>
 +
<tr>
 +
<td> Cutsmart </td>
 +
<td> 1.0 </td>
 +
</tr>
 +
<tr>
 +
<td> XbaI </td>
 +
<td> 0.5 </td>
 +
</tr>
 +
<tr>
 +
<td> DNA </td>
 +
<td> 1.0 </td>
 +
</tr>
 +
</table>
 +
</div>
 +
</div>
 +
 +
<div class="column half_size">
 +
<div align="center">
 +
<h5> Digest of CR-1 </h5>
 +
<table>
 +
<tr>
 +
<th> Content </th>
 +
<th> Experimental ul </th>
 +
<th> Control ul </th>
 +
</tr>
 +
<tr>
 +
<td> H<sub>2</sub>O </td>
 +
<td> 5.0 </td>
 +
<td> 6.0 </td>
 +
</tr>
 +
<tr>
 +
<td> Cutsmart </td>
 +
<td> 1.0 </td>
 +
<td> 1.0 </td>
 +
</tr>
 +
<tr>
 +
<td> EcoRI-HF </td>
 +
<td> 0.5 </td>
 +
<td> X </td>
 +
</tr>
 +
<tr>
 +
<td> XbaI </td>
 +
<td> 0.5 </td>
 +
<td> X </td>
 +
</tr>
 +
<tr>
 +
<td> DNA </td>
 +
<td> 3 </td>
 +
<td> 3 </td>
 +
</tr>
 +
</table>
 +
</div>
 +
</div>
 +
 +
<div class="column half_size">
 +
<div align="center">
 +
<h5> Digest of Dean Vector #3 </h5>
 +
<table>
 +
<tr>
 +
<th> Content </th>
 +
<th> Experimental ul </th>
 +
<th> Control ul </th>
 +
</tr>
 +
<tr>
 +
<td> H<sub>2</sub>O </td>
 +
<td> 2.0 </td>
 +
<td> 3.0 </td>
 +
</tr>
 +
<tr>
 +
<td> Cutsmart </td>
 +
<td> 1.0 </td>
 +
<td> 1.0 </td>
 +
</tr>
 +
<tr>
 +
<td> EcoRI-HF </td>
 +
<td> 0.5 </td>
 +
<td> X </td>
 +
</tr>
 +
<tr>
 +
<td> XhoI </td>
 +
<td> 0.5 </td>
 +
<td> X </td>
 +
</tr>
 +
<tr>
 +
<td> Vector </td>
 +
<td> 6 </td>
 +
<td> 6 </td>
 +
</tr>
 +
</table>
 +
</div>
 +
</div>
 +
 +
<div class="column half_size">
 +
<div align="center">
 +
<h5> PCR </h5>
 +
<table>
 +
<tr>
 +
<th> Contents </th>
 +
<th> Trial 1 (ul) </th>
 +
<th> Trial 2 (ul) </th>
 +
</tr>
 +
<td> Phire </td>
 +
<td> 25.0 </td>
 +
<td> 25.0 </td>
 +
<td> 25.0 </td>
 +
</tr>
 +
<tr>
 +
<td> H<sub>2</sub>O</td>
 +
<td> 21.5 </td>
 +
<td> 19.0 </td>
 +
<td> 14.0 </td>
 +
</tr>
 +
<tr>
 +
<td> 10 uM F Primer </td>
 +
<td> 1.25 </td>
 +
<td> 1.50 </td>
 +
<td> 5.0  </td>
 +
</tr>
 +
<tr>
 +
<td> 10 uM R Primer </td>
 +
<td> 1.25 </td>
 +
<td> 1.50 </td>
 +
<td> 5.0
 +
</tr>
 +
<tr>
 +
<td> 100pg/ul Construct </td>
 +
<td> 1 </td>
 +
<td> 1 </td>
 +
<td> 1 </td>
 +
</tr>
 +
</table>
 +
</div>
 +
</div>
 +
 +
<div class="column half_size">
 +
<div align="center">
 +
<h5> PCR Settings: 30 cycles </h5>
 +
<table>
 +
<tr>
 +
<th> Phase </th>
 +
<th> Temperature (°C) </th>
 +
<th> Time (sec)</th>
 +
</tr>
 +
<tr>
 +
<td> Initial Denaturation </td>
 +
<td> 98 </td>
 +
<td> 30 </td>
 +
</tr>
 +
<tr>
 +
<td> Denaturation </td>
 +
<td> 98 </td>
 +
<td> 5 </td>
 +
</tr>
 +
<tr>
 +
<td> Annealing </td>
 +
<td> 66 </td>
 +
<td> 10 </td>
 +
</tr>
 +
<tr>
 +
<td> Extension </td>
 +
<td> 72 </td>
 +
<td> 20 </td>
 +
</tr>
 +
<tr>
 +
<td> Final Extension </td>
 +
<td> 72 </td>
 +
<td> 60 </td>
 +
</tr>
 +
</table>
 
</div>
 
</div>
 
</div>
 
</div>

Revision as of 02:21, 7 September 2016

Week 11: 9/5 - 9/11

Week 11: 9/5 - 9/11



9/5

Digest of CR-1 Plasmids

Content Experimental ul Control ul
H2O 15.5 16.5
Cutsmart 2.5 2.5
XhoI 0.5 X
EcoRI-HF 0.5 X
DNA 6 6


PCR of Toggle 2

PCR
Contents Trial 1 (ul) Trial 2 (ul)
Q5 MasterMix 12.5 12.5
H2O 6.5 5.5
10 uM F Primer 0.5 1.0
10 uM R Primer 0.5 1.0
100pg/ul Construct 5 5
PCR Settings: 30 cycles
Phase Temperature (°C) Time (sec)
Initial Denaturation 98 30
Denaturation 98 10
Annealing 68 20
Extension 72 60
Final Extension 72 120

Ran Digests


Digest of J6100
Content Experimental ul
H2O 7.5
Cutsmart 1.0
XbaI 0.5
DNA 1.0
Digest of CR-1
Content Experimental ul Control ul
H2O 5.0 6.0
Cutsmart 1.0 1.0
EcoRI-HF 0.5 X
XbaI 0.5 X
DNA 3 3
Digest of Dean Vector #3
Content Experimental ul Control ul
H2O 2.0 3.0
Cutsmart 1.0 1.0
EcoRI-HF 0.5 X
XhoI 0.5 X
Vector 6 6
PCR
Contents Trial 1 (ul) Trial 2 (ul)
Phire 25.0 25.0 25.0
H2O 21.5 19.0 14.0
10 uM F Primer 1.25 1.50 5.0
10 uM R Primer 1.25 1.50 5.0
100pg/ul Construct 1 1 1
PCR Settings: 30 cycles
Phase Temperature (°C) Time (sec)
Initial Denaturation 98 30
Denaturation 98 5
Annealing 66 10
Extension 72 20
Final Extension 72 60


9/6



9/7



9/8



9/9



9/10



9/11