Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2014/11/06

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mm/dd/yyyy

  • Retried PCR of golden gate parts using Rene's primers, this time doing a Tm screen
Reagent uL
ddH2O 33
HF buffer 10
primer F (10uM) 2.5
primer R (10uM) 2.5
dNTP (10mM) 1
Template K084007 0.5
Phusion polymerase 0.5
Total 50



  • annealed for 30 seconds
  • used Tm: 30, 32, 34, 36, 38
Number Template F Primer R Primer Annealing T Expected length
1 K084007 3/10/13 4-2-1 P069 P070 32 645
2 K084007 3/10/13 4-2-2 P069 P070 32 645



  • Still no clear bands observed for any of the annealing temperatures.