User:Karmella Haynes/Notebook/BioBrick cloning/2012/07/26

From OpenWetWare
Jump to: navigation, search
Owwnotebook icon.pngKarmella's BioBrick Cloning Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

07/26/12

  • ✓ Sequencing order: DNASU
  • ✓ Minipreps digests: Francesca's plasmids and V0120



Sequencing order

Measure concentrations:

Sample 260 260/280 ng/μL
FCt1 0.095 1.928 94.595
FCt2 0.174 1.903 173.76
FCt4 0.068 1.937 68.042


> DNASU @Biodesign, Results:

  1. FCt1 - VF2: good, P2547 confirmed
  2. FCt2 - VF2: bad seq, wrong primer (Francesca: part is in pGA18, not pSB1A2)
  3. FCt4 - VF2: good, but operator is O1, not O2

> Need to re-do FCt2 (use M13 primer) to find out if it's O2 or actually O1



Minipreps
> Note: Streaks for FCt1 and FCt2 are red; FCt4 is very pale pink
> Check Francesca's plasmids with S/P digests; should remove RBS-RFP (703 bp); see http://partsregistry.org/Part:BBa_J61002
> Check ccdB-V0120 with E/P digests


Reagent Volume Expected:
1. FCt1 (P2547) = ~2250, 703
2. FCt2 (O2-RFP/pGA18) = ~3000, 703
3. FCt4 (P2547-O2-RFP) = ~2250, 703
4-6. V0120 = ~3200, ~1700
Hover name
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 2.0 μL
10X buffer 1.5
enzyme 1 1.0
enzyme 2 1.0
dH2O 9.5
  15 μL --> 37°C/ ~15 min.