Haynes Lab:Notebook/David Barclay Undergrad Training/Plasmid Transformation/2013/09/13: Difference between revisions

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| bgcolor=#cfcfcf | Volume
| bgcolor=#cfcfcf | Volume
| rowspan="7" | <u>Expected:</u><br>1. hPCD = 186 bp, 4968 bp<br>2. Ubc = 368 bp, 4968 bp<br>
| rowspan="7" | <u>Expected:</u><br>1. hPCD = 186 bp, 4968 bp<br>2. Ubc = 368 bp, 4968 bp<br>
| rowspan="7" | [[Image:IMAG0329.jpg|400px|Today's gel]]<br>15 μL/lane; 1% agarose
| rowspan="7" | [[Image:IMAG0329.jpg|400px|Today's gel]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
|-
|-
| DNA(plasmid) || 3.0 μL
| DNA(plasmid) || 3.0 μL

Revision as of 16:27, 13 September 2013

Haynes BioBrick Cloning <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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Day 3- Minipreps

Liquid Culture Results

NOTE: Cultures and Streaks taken out after 11 hours (37 incubator) and left on lab bench for 11 hours.

hPCD

  • Cultures Innoculated 09/12/13
  • Cultures 1 and 2: Both show growth
  • Streak 1 and 2: Both show growth

Ubc

  • Cultures Innoculated 09/12/13
  • Cultures 1 and 2: Both show growth
  • Streak 1 and 2: Both show growth

Samples for Minipreps and Analysis

  • Sample 1= hPCD/V0120 colony 1; BBa_J176000; part= 186 bp; vector= 4968 bp
  • Sample 2= hPCD/V0120 colony 2; same
  • Sample 3= Ubc/V0120 colony 1; BBa_J176003; part= 368 bp; vector= 4968 bp
  • Sample 4= Ubc/V0120 colony 2; same
  • Sample 5= hPCD/V0120; BBa_J176000; part= 186 bp; vector= 4968 bp
  • Sample 6= Ubc/V0120; BBa_J176003; part= 368 bp; vector= 4968 bp

DNA Concentration Data

Plasmid OD260 OD260/280 ng/μL
1. hPCD 1 0.279 1.831 278.676
2. hPCD 2 0.242 1.834 241.754
3. Ubc 1 0.097 1.787 96.614
4. Ubc 2 0.146 1.789 145.548


Restriction Digest Table

  • Checked plasmid minipreps with EcoRI/PstI digests
Reagent Volume Expected:
1. hPCD = 186 bp, 4968 bp
2. Ubc = 368 bp, 4968 bp
 Today's gel
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 3.0 μL
10X FD 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 P001
  2. Plasmid 1 - Reverse P002
  3. Plasmid 2 - Forward P001
  4. Plasmid 2 - Reverse P002
  5. Plasmid 3 - Forward P001
  6. Plasmid 3 - Reverse P002
  7. Plasmid 4 - Forward P001
  8. Plasmid 4 - Reverse P002


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