User:Karmella Haynes/Notebook/BioBrick cloning/2013/09/06

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09/06/13

  • Miniprep - CMV
  • Restriction digest - CMV
  • Sequencing - CMV [cloning bootcamp demo]


Results

  • CMV/V0120
    • Culture innoculated on 9/05/13
    • Results for liquid cultures: (1) grew, (2) grew
    • Results for streak plate: (1) grew well, (2) hardly any growth


List of Samples for Minprep & Analysis

  1. Sample 1 = CMV/V0120 colony 1; BBa_J176027; part = 588 bp; vector = 3200
  2. Sample 2 = CMV/V0120 colony 2; same
  3. Sample 3 = CMV/V0120 (control DNA); BBa_J176027; part = 588 bp; vector = 3200


DNA Concentration Data

Plasmid OD260 OD260/280 ng/μL
1. Plasmid 1 0.108 1.841 108.321
2. Plasmid 2 0.007 1.75 6.60
  • Streak #2 did not grow well. Culture might be empty bacteria. Use mini prep 1 for other steps.


Restriction Digest Table

  • Checked plasmid minipreps with EcoRI/PstI digests
Reagent Volume Expected:
1. Plasmid 1 = insert size, vector size
2. Plasmid 2 = insert size, vector size
 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 'date'
  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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