Difference between revisions of "Team:Stony Brook/Notebook/Cancer-W6"

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<h6> Emmanuel Ligation (193 ng/ul) </h6>
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<h4> Emmanuel Ligation (193 ng/ul) </h4>
 
<ul>
 
<ul>
 
<table>
 
<table>
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<br>
 
<br>
  
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<h6> Gel run </h6>
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<h4> Gel run </h4>
 
<p> Lanes: </p>
 
<p> Lanes: </p>
 
<ol>
 
<ol>
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<br>  
  
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<h6> 25 ul Digest of Ryan + Emmanuel (Veronica) Miniprep </h6>
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<h4> 25 ul Digest of Ryan + Emmanuel (Veronica) Miniprep </h4>
 
<ul>
 
<ul>
 
<li> Made 17 LB With Ampicillin (600ml)</li>
 
<li> Made 17 LB With Ampicillin (600ml)</li>
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<h6> Miniprep </h6>
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<h4> Miniprep </h4>
 
<ul>  
 
<ul>  
 
<li> Miniprep of ligated construct </li>  
 
<li> Miniprep of ligated construct </li>  
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</ul>
  
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<h6> Ran a nanodrop of Minipreps </h6>
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<h4> Ran a nanodrop of Minipreps </h4>
 
<ul>
 
<ul>
 
<table>
 
<table>
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</ul>
 
</ul>
  
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<h6> Ran a gel of miniprepped plasmids </h6>
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<h4> Ran a gel of miniprepped plasmids </h4>
 
<ul>
 
<ul>
 
<li> Used 5ul of DNA and 1 ul of dye </li>
 
<li> Used 5ul of DNA and 1 ul of dye </li>
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<h6> Digest of Miniprepped Plasmids </h6>
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<h4> Digest of Miniprepped Plasmids </h4>
 
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<h6> Transformations </h6>
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<h4> Transformations </h4>
 
<ul>  
 
<ul>  
 
<li> Transformed R001, J61100, puC19 </li>  
 
<li> Transformed R001, J61100, puC19 </li>  
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</ul>
 
</ul>
  
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<h6>Miniprep of BioBricks from 8/2</h6>
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<h4>Miniprep of BioBricks from 8/2</h4>
  
−
<h6> Gel run ( 6uL of each tube loaded)</h6>
+
<h4> Gel run ( 6uL of each tube loaded)</h4>
 
<p> Lanes: </p>
 
<p> Lanes: </p>
 
<ol>
 
<ol>
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</ul>
 
</ul>
  
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<h6>Interlab LB cultures made from plates </h6>
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<h4>Interlab LB cultures made from plates </h4>
  
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<h6> Developed sequencing primers for construct </h6>
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<h4> Developed sequencing primers for construct </h4>
  
 
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<div class="column full_size">
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<h6>Worked on InterLab study </h6>
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<h4>Worked on InterLab study </h4>
  
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<h6> Gel ran (done by Emmanuel) </h6>
+
<h4> Gel ran (done by Emmanuel) </h4>
 
<p> Lanes: </p>
 
<p> Lanes: </p>
 
<ol>
 
<ol>

Revision as of 20:31, 4 August 2016

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Week 4: 8/1-8/5

8/1-8/9

8/1

Emmanuel Ligation (193 ng/ul)

    Content Experimental (ul) Control (ul)
    H2O 38.8 39.8
    Cutsmart 5.0 5.0
    XbaI 0.5 X
    EcoRI-HF 0.5 X
    DNA 5.18 5.18

Gel run

Lanes:

  1. Ladder
  2. Emmanuel Digest Control
  3. Emmanuel Digest Experimental
  • Did not work

25 ul Digest of Ryan + Emmanuel (Veronica) Miniprep

  • Made 17 LB With Ampicillin (600ml)
Ryan - 446.4 ng/ul
    Content Experimental (ul) Control (ul)
    H2O 20.4 21.4
    Cutsmart 2.5 2.5
    XbaI 0.5 X
    EcoRI-HF 0.5 X
    DNA 1.12 1.12
Emmanuel - 193 ng/ul
    Content Experimental (ul) Control (ul)
    H2O 18.9 19.9
    Cutsmart 2.5 2.5
    XbaI 0.5 X
    EcoRI-HF 0.5 X
    DNA 2.59 2.59

8/2

Miniprep

  • Miniprep of ligated construct
  • 10,000rpm for 3 minutes at 25°C

Ran a nanodrop of Minipreps

    Content Concentration (ng/ul) 260/280
    Veronica 1 311.5 1.86
    Veronica 2 43.8 1.76
    Veronica 3 259 1.88
    Veronica 4 296 1.88
    R1 176.5 1.75
    Ryan 2 277.3 1.85
    Ryan 3 301.6 1.81
    Ryan 4 366.6 1.82

Ran a gel of miniprepped plasmids

  • Used 5ul of DNA and 1 ul of dye

Lanes:

  1. Veronica 1
  2. Veronica 3
  3. Veronica 4
  4. Ladder
  5. Ryan 1
  6. Ryan 2
  7. Ryan 3
  8. Ryan 4

Digest of Miniprepped Plasmids

Veronica 1 - 311.5 ng/ul
    Content Experimental (ul) Control (ul)
    H2O 40.8 41.8
    Cutsmart 5 5
    XbaI 0.5 X
    PvuII-HF 0.5 X
    DNA 3.21 3.21
Ryan 4 - 366.6 ng/ul
    Content Experimental (ul) Control (ul)
    H2O 41.3 42.3
    Cutsmart 5 5
    XbaI 0.5 X
    PvuII-HF 0.5 X
    DNA 2.73 2.73

8/3

Transformations

  • Transformed R001, J61100, puC19
  • 500 uL of SOC used for each transformation

Miniprep of BioBricks from 8/2

Gel run ( 6uL of each tube loaded)

Lanes:

  1. 5 uL plasmid
  2. 5 uL plasmid + 5 uL of nuclease free water
  3. 5 uL plasmid + 5 uL water + 2.5 uL of CutSmart Buffer
  4. Ladder
  5. 5 uL plasmid + 5 uL water + 2.5 uL CutSmart + 0.75 uL XhoI
  6. 5 ul plasmid + 5 uL water + 2.5 uL CutSmart + 0.75 uL PvuII
  7. 5 ul plasmid + 6 uL water + 2.5 uL CutSmart + 0.75 uL XhoI + 0.75 uL PvuII
  • Did not work

Interlab LB cultures made from plates

Developed sequencing primers for construct

8/4

Worked on InterLab study

Gel ran (done by Emmanuel)

Lanes:

  1. Veronica control (plasmid)
  2. Veronica digest
  3. Ladder
  4. Ryan digest
  5. Ryan control (plasmid)
  • Did not work