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Homemade qPCR Mix Recipe

  • 10x Homemade Core PCR Buffer 100mM Tris HCl (pH 8.5)
    • 200mM KCl
    • 30mM MgCl2
    • 1.5% Triton X-100
  • 500nM Fluorescein Passive Reference Dye (USB, Product code: 75767)
  • SYBR Green dye (Invitrogen, Product code: S-7563)) diluted 1:100 in  dimethyl sulfoxide (DMSO) (Sigma, Product code: D2438)
  • BSA (bovine serum albumin) (Biolabs, Product code: B9001S )
  • Formamide (BDH Lab. Supplies, Product code: 103266T)
For a 20µl reaction: Component  Volume (µl)  Final Concentration
10x homemade core buffer21x
2mM dNTP mix20.2mM
20µM primer mix0.5500nM
500nM Fluorescein Dye0.410nM
10mg/ml BSA0.40.2mg/ml
0.01 SYBR green dye0.15.0e-5
Taq (Bioline; 5U/ul)0.21 Units

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