User:Morgan L. Paull/Notebook/Genetic Fence/2011/01/17

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Diagnostic Digest of GalUAS35s parts

  • Cut the two tubes I have each with xba pst and eco pst. The two tubes were an unlabeled one and one labeled as a ligation mix.
  • All reactions were 10μL, with 1μL FD Green buffer
  • Used 7μL of the gel extracted V0120 Backbone from yesterday, and 1μL of the 35s annealed oligos, at the concentrations listed below


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KB+ Ladder 5xGalUAS (unlabeled) Eco Pst 5xGalUAS (ligation) Eco Pst 5xGalUAS (unlabeled) Xba Pst 5xGalUAS (ligation) Xba Pst Ligation mix plasmid Unlabeled plasmid KB+

Ligating 35s.A.2 anneal into V0120

1x insert concentration

  • 1μL 35s
  • 7μL BB
  • 2μL #3 Roche buffer
  • 10μL #4 Roche buffer
  • 1μL #5 Roche buffer

1/10x insert concentration

  • 1μL 1/10 35s
  • 7μL BB
  • 2μL #3 Roche buffer
  • 10μL #4 Roche buffer
  • 1μL #5 Roche buffer

1/100x insert concentration

  • 1μL 1/100 35s
  • 7μL BB
  • 2μL #3 Roche buffer
  • 10μL #4 Roche buffer
  • 1μL #5 Roche buffer

1/1000x insert concentration

  • 1μL 1/1000 35s
  • 7μL BB
  • 2μL #3 Roche buffer
  • 10μL #4 Roche buffer
  • 1μL #5 Roche buffer

Control

  • 1μL DH2O
  • 7μL BB
  • 2μL #3 Roche buffer
  • 10μL #4 Roche buffer
  • 1μL #5 Roche buffer


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