Haynes Lab:Notebook/Jan/2013/09/27

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(Plasmid Extraction and Validation of fshPCD)
(Plasmid Extraction and Validation of fshPCD)
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| bgcolor=#cfcfcf | Reagent  
| bgcolor=#cfcfcf | Reagent  
| bgcolor=#cfcfcf | Volume
| bgcolor=#cfcfcf | Volume
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| rowspan="7" | <u>Expected:</u><br>1. Plasmid 1 = insert size, vector size<br>2. Plasmid 2 = insert size, vector size<br>
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| rowspan="7" | <u>Expected:</u><br>1. Plasmid 1 = 186 bp, 4986 <br>2. Plasmid 2 = 186, 4986 <br>
| rowspan="7" | [[Image:GelImage.jpg|400px|Today's gel]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
| rowspan="7" | [[Image:GelImage.jpg|400px|Today's gel]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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Revision as of 19:20, 4 October 2013

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Plasmid Extraction and Validation of fshPCD

  • Name: fshPCD / BBa_J176002
    • Date of culture inoculation: 9/26/13
    • Results for Cultures 1 and 2: Mixture is cloudy so growth in both. Success.
    • Results for Streaks 1 and 2: Growth in both areas along zig-zag line. Bottom left of agar plate dried out. Success.
  • Sample 1: fshPCD/V0120 colony 1; BBa_J176002; part = 186 bp; vector = 4986 bp
  • Sample 2: fshPCD/V0120 colony 2; same
  • Sample 3: fshPCD/V0120 (control DNA); BBa_J176002; part = 186 bp; vector = 4986 bp

DNA Concentration Data

Plasmid OD260 OD260/280 ng/μL
1. Plasmid 1 0.186 1.859 185.784
2. Plasmid 2 0.138 1.827 138.008


Restriction Digest Table

  • Checked plasmid minipreps with EcoRI/PstI digests
Reagent Volume Expected:
1. Plasmid 1 = 186 bp, 4986
2. Plasmid 2 = 186, 4986
Image:GelImage.jpg
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 3.0 μL
10X buffer 1.5
EcoRI 1.0
PstI 1.0
dH2O 8.5
  15 μL --> 37°C/ 15 min.


DNA Sequencing Samples

  • Submitted to DNASU on: 'date'
  • Order Number:
  1. Plasmid 1 - forward primer 'name'
  2. Plasmid 1 - reverse primer 'name'
  3. Plasmid 2 - forward primer 'name'
  4. Plasmid 2 - reverse primer 'name'




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