Difference between revisions of "Team:Stony Brook/Notebook/Cancer-W10"

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<h4> Digest of IDT CR-1 SBU and Vector  </h4>
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</div>
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<div align="center">
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<h5> CR-1 SBU  </h5>
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<ul>
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<table>
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<tr>
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<th> Content </th>
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<th> Experimental ul </th>
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<th> Control ul </th>
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</tr>
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<tr>
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<td> H<sub>2</sub>O </td>
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<td> 18</td>
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<td> 20 </td>
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</tr>
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<tr>
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<td> Cutsmart </td>
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<td> 5.0 </td>
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<td> 5.0 </td>
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</tr>
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<tr>
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<td> EcoRI-HF </td>
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<td> 1.0 </td>
 +
<td> X </td>
 +
</tr>
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<tr>
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<td> XhoI </td>
 +
<td> 1.0 </td>
 +
<td> X </td>
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</tr>
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<tr>
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<td> DNA </td>
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<td> 25.0 </td>
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<td> 25.0 </td>
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</tr>
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</table>
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</ul>
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</div>
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<div class="column half_size">
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<div align="center">
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<h5> pEP352GAP </h5>
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</div>
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<ul>
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<table>
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<tr>
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<th> Content </th>
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<th> Experimental ul </th>
 +
<th> Control ul </th>
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</tr>
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<tr>
 +
<td> H<sub>2</sub>O </td>
 +
<td> 7.0 </td>
 +
<td> 8.0 </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> DNA </td>
 +
<td> 1.0 </td>
 +
<td> 1.0 </td>
 +
</tr>
 +
</table>
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</ul>
 +
</div>
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<div align="center">
 
<div align="center">

Revision as of 03:12, 7 September 2016

Week 10: 8/29 - 9/4

Week 10: 8/29 - 9/4



8/29

AmilCP BioBrick Digests

Content Experimental ul Control ul
H2O 46.5 47.0
Cutsmart 5.0 5.0
XbaI 0.5 X
DNA 2.0 2.0

PCR of Toggle 1

Contents ul
Phire 10
H2O 8
5 uM F Primer 1.5
5 uM R Primer 1.5
100pg/ul Construct 1.0

  • Ran other trials with 2.5, 3.5, 4.5 and 6.5 ul of Forward and Reverse Primers
  • Water ul was adjusted to compensate: 6, 4, 2 and 0 ul respectively
Phase Temperature (°C) Time (sec)
Initial Denaturation 98 30
Denaturation 98 5
Annealing 68 5
Extension 72 45
Final Extension 72 60

PCR of Toggle 2

Contents ul
Phire 10
H2O 8
5 uM F Primer 1.5
5 uM R Primer 1.5
100pg/ul Construct 1.0

  • Ran other trials with 2.5, 3.5, 4.5 and 6.5 ul of Forward and Reverse Primers
  • Water ul was adjusted to compensate: 6, 4, 2 and 0 ul respectively
Phase Temperature (°C) Time (sec)
Initial Denaturation 98 30
Denaturation 98 5
Annealing 68 5
Extension 72 45
Final Extension 72 60

Digests of Toggle parts

Toggle 1 Digest
Content Experimental ul Control ul
H2O 24 25
Cutsmart 5.0 5.0
SpeI 1.0 X
DNA 25.0 25.0
Toggle 1 Digest
Content Experimental ul Control ul
H2O 24 25
Cutsmart 5.0 5.0
XbaI 1.0 X
DNA 25.0 25.0

Linearized pSB1C7 Digest


Content Experimental ul Control ul
H2O 16.5 18.5
Cutsmart 2.5 2.5
SpeI-HF 1.0 X
XbaI 1.0 X
DNA 4.0 4.0


Ligation of Toggle parts

Content ul
10X T4 Buffer 2.0
Toggle 1 5.0
Toggle 2 10.0
H2O 2.0
Ligase 1.0


8/30

Ligate Toggle 2 into Plasmid

    Content ul
    10X T4 Buffer
    Vector # (# ng)
    Construct # (# ng)
    H2O #
    Ligase #


8/31

Digest of IDT CR-1 SBU and Vector

CR-1 SBU
    Content Experimental ul Control ul
    H2O 18 20
    Cutsmart 5.0 5.0
    EcoRI-HF 1.0 X
    XhoI 1.0 X
    DNA 25.0 25.0
pEP352GAP
    Content Experimental ul Control ul
    H2O 7.0 8.0
    Cutsmart 1.0 1.0
    EcoRI-HF 0.5 X
    XhoI 0.5 X
    DNA 1.0 1.0


9/1



9/2



9/3



9/4