User:Robwarden/Notebook/Caspase-3 Activatable Probe/2009/09/04

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Primer Design for hRLuc8

  • hRLuc8_EcoRI_Fwd
    • 5'-gATCgAATTCTATggCTTCCAAggTgTACgAC-3'
    • 5'-Spacer-EcoRI-Frame Correction-Annealing-3'
    • Tm = 64, 58 (annealing)
    • %GC = 47, 52 (annealing)
  • hRLuc8_XhoI_Rev
    • 5'-gATCCTCgAgCTgCTCgTTCTTCAgCACg-3'
    • 5'-Spacer-XhoI-Annealing-3'
    • Tm = 67, 57 (annealing)
    • %GC = 59, 58 (annealing

PCR of hgLuc

Reaction Setup

 5.0 μL 10X Buffer
 1.0 μL 1:20 dil'n hgLuc plasmid
 2.5 μL Primer "hgLuc Rev XhoI"
 2.5 μL Primer "hgLuc For EcoRI"
 1.0 μL 10mM dNTP mix
 0.5 μL Vent Polymerase
37.5 μL dH2O

Thermo Cycle

Cycles Temp (°C) Time
1 94 10:00
30 94 0:45
50 0:30
72 1:30
1 72 10:00
4



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