Haynes Lab:Notebook/SynBERC Scholars 2013/2014/03/18

From OpenWetWare

Jump to: navigation, search
Project name Main project page
Previous entry      

03/18/14

  • Dilution-limited PCR: Kozak plasmid and Luc14 genomic DNA



Dilution-limited PCR

  • K = Kozak plasmid DNA, initial concentration = 243 ng/μL
  • L = Luc 14 genomic DNA, initial concentration = 217 ng/μL


Dilutions for PCR Reactions

  • K1 = 10-3, 120.0 pg/ 0.5 μL
  • K2 = 10-6, 0.120 pg/ 0.5 μL
  • K3 = 10-9, 120.0 fg/ 0.5 μL
  • K4 = 10-12, 0.120 fg/ 0.5 μL
  • L1, L5 = 10-1, 10.8 ng/ 0.5 μL
  • L2, L6 = 10-2, 1.108 ng/ 0.5 μL
  • L3, L7 = 10-3, 108 pg/ 0.5 μL
  • L4, L8 = 10-4, 10.8 pg/ 0.5 μL


PCR Reactions

  • Same conditions as optimization (3/17/14), except ran 40 cycles instead of 35
  • 96 reactions total


Fluorimeter measurements

  • Added 125 dH2O to each PCR reaction to dilute DNA 1:6
  • Diluted SYBR Green 1 (Promega) to 10x working concentration in 1x TBE



Personal tools