2×SYBR qPCR Mix
This ready‑to‑use kit is designed for fluorescence‑based real‑time quantitative PCR (qPCR) of target fragments. It contains all essential qPCR components: optimized buffer, Taq DNA polymerase, dNTPs, MgCl₂ and fluorescent dye. Users only add templates and primers to run qPCR and HRM analysis for versatile applications.
Product Features
- Simple workflow: only template and primers are needed for qPCR setup.
- Adopts third‑generation saturated fluorescent dye with strong signal and no PCR inhibition.
- Pre‑formulated premix simplifies pipetting, reduces cross‑contamination and experimental errors.
- Carefully balanced components deliver high sensitivity and specificity; detection limit as low as 50 copies/mL.
Package Specification
Storage
Store at -20 ℃. Shelf life: 6 months.
Reference 20 μL qPCR Reaction System
| Component |
Volume for 20 μL system |
Final Concentration |
| 2× SYBR qPCR Mix |
10 μL |
1 × |
| Forward Primer |
variable |
0.1–0.5 μM |
| Reverse Primer |
variable |
0.1–0.5 μM |
| Template |
variable |
— |
| ROX (user-supplied) |
2 μL, instrument-dependent |
— |
| Ultrapure water |
to 20 μL |
— |
Load the plate into a real‑time PCR instrument and collect fluorescence signals at annealing or extension stage.
Notes
A primer final concentration of 0.2 μM usually gives good results; adjustable range: 0.1–1.0 μM.
Recommended template input: 10–100 ng genomic DNA or 1–10 ng cDNA. Since target copy numbers vary by sample type, template gradient dilution is recommended to find the optimal loading amount.
ROX Passive Reference Dye Guidance
ROX is required for Ct correction on ABI 7500, 7700 and 7900 systems.
- ABI 7700 / 7900: 1× ROX; add 2 μL of 10× ROX per 20 μL reaction.
- ABI 7500: 0.05–0.1× ROX; add 0.1–0.2 μL 10× ROX per 20 μL reaction, or dilute 10× ROX to 1× and add 1–2 μL.
ROX may introduce background noise on melting curves and cause false peaks. If spurious peaks appear, disable ROX in the Passive Reference Dye setting of your software and re-analyze.
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