Haynes Lab:Notebook/Jan/2013/10/18

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(Extraction and Verification of mCh)
(Extraction and Verification of mCh)
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| bgcolor=#cfcfcf | Volume
| bgcolor=#cfcfcf | Volume
| rowspan="7" | <u>Expected:</u><br>1. Plasmid 1 = 705 bp, 3200 bp<br>2. Plasmid 2 = 705 bp, 3200 bp<br>
| rowspan="7" | <u>Expected:</u><br>1. Plasmid 1 = 705 bp, 3200 bp<br>2. Plasmid 2 = 705 bp, 3200 bp<br>
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| 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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| rowspan="7" | [[Image:Jan_Simper_mCh_DNA_Boot_Camp.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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| DNA(plasmid) || 3.0 μL
| DNA(plasmid) || 3.0 μL

Revision as of 15:12, 9 November 2013

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Extraction and Verification of mCh

  • Name: mCh / BBa_J176005
    • Date of culture inoculation: 10/17/13
    • 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, although more growth in colony 1 streak than colony 2 streak. Overall success.
  • Sample 1: mCh/V0120 colony 1; BBa_J176005; part = 705 bp; vector = 3200 bp
  • Sample 2: mCh/V0120 colony 2; same
  • Sample 3: mCh/V0120 (control DNA); BBa_J176005; part = 705 bp; vector = 3200 bp


DNA Concentration Data

Plasmid OD260 OD260/280 ng/μL
1. Plasmid 1 0.12 1.84 120
2. Plasmid 2 0.026 1.656 26


Restriction Digest Table

  • Checked plasmid minipreps with EcoRI/PstI digests
Reagent Volume Expected:
1. Plasmid 1 = 705 bp, 3200 bp
2. Plasmid 2 = 705 bp, 3200 bp
Today's gel
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: '10/21/13'
  • Order Number: 8258
  1. mCh - forward primer 'P0001' -- 705 matches, thus 100% matching across the sequence. DNA cloning successful.
  2. mCh - reverse primer 'P0002' -- 705 matches, so 100% match. DNA cloning successful.



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