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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 |
| H2O | 12.9 | |
| 10x homemade core buffer | 2 | 1x |
| 2mM dNTP mix | 2 | 0.2mM |
| 20µM primer mix | 0.5 | 500nM |
| 100%formamide | 0.5 | 2.5% |
| 500nM Fluorescein Dye | 0.4 | 10nM |
| 10mg/ml BSA | 0.4 | 0.2mg/ml |
| 0.01 SYBR green dye | 0.1 | 5.0e-5 |
| Taq (Bioline; 5U/ul) | 0.2 | 1 Units |
| Template | 1 |
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