Haynes Lab:Notebook/Engineering PC-TFs/2012/02/16

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Ligation/Assembly

  • Weight gel slices.
  • Add three times amount of volume to each tube of gel (600 µL)
  • Incubate at 55°C for 5-10 minutes.
  • Centrifuge for a minute at 17,000 x g.
  • Add 20 µL of wash buffer.
  • Don't dump supernatent and centrifuge for one minute again.
  • Place spin columns in 1.5 mL tubes.

Measure concentration of each fragment:

  • Load Gen 5 2.0 software. Go to the Task Manager-Take 3 Application
  • Go to Nucleic Acid Quantification.
  • Pipette 2 µL of each DNA sequence into Take 3 plate.
  • Place plate in EPOCH plate reader.

Results: 1.93 260 1.069 280 1.805 260/280 96.477 ng/µL 2.443 260 1.18 280 2.069 260/280 122.149 ng/µL 1.604 260 0.831 280 1.929 260/280 80.182 ng/µL 0.541 260 0.258 280 2.094 260/280 27.054 ng/µL 0.942 260 0.468 280 2.013 260/280 47.105 ng/µL 0.904 260 0.434 280 2.083 260/280 45.217 ng/µL 1.321 260 0.655 280 2.015 260/280 66.047 ng/µL



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