SynBioBootcamp:Notebook/SynBioBootcamp09:Notebook/SynBioBootcamp09

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Notes

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  • Oligos (2/18/09)
  • 1) Performed quick spin on tubes to pellet the DNA powder
  • 2) Added 288 uL ddH20 to 288 nmol of oligos to a final concentration of 100uM
  • 3) Took out a 1uL aliquot and diluted 10X to 10 10uM fpr PCR
  • PCR (2/18/09)
  • 1) Prepared PCR reaction for pAC_Tet_inv-short using SBB-Protocol PCR2
  • 2) Added 0.5 expand polymerase "1"
  • 3) Used the C4K55 thermocycler ptotocol for an expected product of ~2.7 kb
  • Analytical Gel (2/23/09)
  • 1) Ran an analytical gel and performed the regular zymo cleanup.
  • 2) Expected 2.7kB band in lane 9 was not observed.
  • 3) Ran a second analytical gel; no expected 2.7 kb band oberved
  • 4) Prepared a new PCR reaction as per 2/18/09 checking off each of the steps in SBB-Protocol PCR2 after each step execution.


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