SBB09 PCR Cloning

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Dilute oligonucleotides to be 10 uM

  • Add 1 ul of oligo dilution to 9 ul of ddH20
  • Mix and spin

Cloning PCR, (products are 937bp & 1600bp, )

  • 24 uL water
  • 3.3 uL Expand 10x Buffer 2
  • 3.3 uL 10x dNTPs (2 mM in each; 0.2 mM final conc)
  • 1 uL Oligo Odvs007F (10uM)
  • 1 uL Oligo Odvs008rR (10uM)
  • 0.5 uL Expand Polymerase 1
  • 0.5 uL Template DNA E.coli 0157:H7
  • Run this PCR on program 55
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