User:Robwarden/Notebook/GFP-RGD Force Sensor/2009/03/10

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GFPst PCR

Primer Dilutions

  • Stock diluted to 100μM
  • Working concentration diluted to 10μM

Reaction

  10 μL 5X HF Buffer
   1 μL 10mM dNTPs
 2.5 μL Forward Primer
 2.5 μL Reverse Primer
   1 μL GFPuv Plasmid (143.4 ng)
 0.5 μL Phusion Hot Start Polymerase
32.5 μL MQ

Thermal Cycle

Cycles Temp (°C) Time
1 98 0:30
35 98 0:10
68 0:30
72 0:30
1 72 5:00
4

Validation

  • 0.8% Agarose Gel w/ EtBr
  • Strong band seen & extracted
  • DNA purified with Qiagen kit