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

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<li> 5 uL plasmid + 5 uL of nuclease free water</li>
 
<li> 5 uL plasmid + 5 uL of nuclease free water</li>
 
<li> 5 uL plasmid + 5 uL water + 2.5 uL of CutSmart Buffer</li>
 
<li> 5 uL plasmid + 5 uL water + 2.5 uL of CutSmart Buffer</li>
 +
<li> Ladder </li>
 
<li> 5 uL plasmid + 5 uL water + 2.5 uL CutSmart + 0.75 uL XhoI </li>
 
<li> 5 uL plasmid + 5 uL water + 2.5 uL CutSmart + 0.75 uL XhoI </li>
 
<li> 5 ul plasmid + 5 uL water + 2.5 uL CutSmart + 0.75 uL PvuII</li>
 
<li> 5 ul plasmid + 5 uL water + 2.5 uL CutSmart + 0.75 uL PvuII</li>
Line 370: Line 371:
  
 
<h6> Developed sequencing primers for construct </h6>
 
<h6> Developed sequencing primers for construct </h6>
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 +
<div class="column full_size">
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<ul>
 +
<hr>
 +
</ul>
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</div>
 +
 +
<div class="column full_size">
 +
<div align="center">
 +
<h2 style="color:#B61C1D;"> 8/4 </h2>
 +
</div>
 +
</div>
 +
 +
<h6>Worked on InterLab study </h6>
 +
 +
<h6> Gel ran (done by Emmanuel) </h6>
 +
<p> Lanes: </p>
 +
<ol>
 +
<li> Veronica control (plasmid) </li>
 +
<li> Veronica digest </li>
 +
<li> Ladder </li>
 +
<li>  Ryan digest</li>
 +
<li> Ryan control (plasmid) </li>
 +
 +
</ol>
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<ul>
 +
<li> Did not work </li>
 +
</ul>

Revision as of 19:37, 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