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ZY Quant Star qPCR Premix
ZY Quant Star qPCR Premix is a ready-to-use 2× master mix containing all components except primers and DNA template.
The antibody-based hot-start enzyme suppresses nonspecific amplification and primer-dimer formation. Universal ROX reference dye provides compatibility with major qPCR platforms.
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  • 2×

    Ready-to-use master mix

  • Universal ROX

    Compatible with major qPCR platforms

  • 80–200 bp

    Recommended amplicon length

  • 40 cycles

    Recommended cycle count

首页 > Products > Molecular Biology Reagents > ZY Quant Star qPCR Premix >
Product Overview
Ordering Information and Key Advantages
Scope of Application
Instructions and Cycling Program
Assay Optimization and Primer Design
Frequently Asked Questions
Related Products and Services

Product Overview

ZY Quant Star qPCR Premix · One-step loading, precise quantification

ZY Quant Star qPCR Premix is a real-time fluorescent quantitative PCR reagent launched by Zhi Yin Biological, specifically designed for the SYBR Green I intercalating dye method. It comes in a 2× premix format, containing all qPCR components except primers and DNA template, significantly simplifying the pipetting process, shortening operation time, and reducing the chance of cross-contamination.

The core component is an antibody-modified hot-start Taq DNA polymerase. Together with an optimized buffer system and PCR reaction enhancers, it can effectively suppress non-specific amplification and primer dimer formation. The premix includes a universal reference dye, compatible with various qPCR instruments that require ROX normalization, without the need for additional dye addition.

  • SYBR Green I dye method
  • Antibody-based hot-start Taq DNA polymerase
  • Universal calibration dye, compatible with mainstream qPCR platforms
ZY Quant Star qPCR Premix real-time fluorescent quantitative PCR

Ordering information

Product Name Catalog No. Size Detection method Price
ZY Quant Star
qPCR Premix 		ZY-QS-1000 1 mL SYBR Green I dye method
(Real-time fluorescent quantitative PCR)
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Core advantages

Ready-to-use, specific, universal, and compatible with high-throughput experiments
  • Ready-to-use 2× premix solution
    Contains all components except primers and template, reducing pipetting steps and contamination risk.
  • Antibody-mediated hot-start enzyme
    Suppresses polymerase activity at low temperature, releases after pre-denaturation, enhancing specificity and sensitivity.
  • Universal calibration dye
    Suitable for validated compatible instruments; for specific models and parameter settings, please refer to the manual.
  • Broad template range
    Maintains good linearity for templates ranging from high to low copy numbers.
  • Easy to operate
    Shortens sample loading time and reduces the chance of system preparation contamination, suitable for high-throughput and repeated experiments.

Scope of Application

qPCR experiments and amplification curves
  • Relative quantification of gene expression (2--ΔΔCt method)
  • Absolute quantification of target gene (standard curve method)
  • Specific quantitative detection of templates such as cDNA and genomic DNA
  • mRNA expression level analysis (combined with RNA extraction and reverse transcription experiments)

Instructions and Cycling Program

Recommended reaction system (10 μL)
Reagents Usage
ZY Quant Star qPCR Premix 5 μL
Forward primer (10 μM) 0.4 μL
Reverse primer (10 μM) 0.4 μL
DNA Template X μL
Nuclease-Free Water Bring up to 10 μL
Two-step reaction program (recommended)
Step Temperature Time Number of cycles
Pre-denaturation 95℃ 30 sec —
Denaturation 95℃ 10 sec 40 cycles
Annealing & Extension 60℃ 30 sec
Melting curve Instrument Default Instrument Default —
Three-Step Reaction Program
Step Temperature Time Number of cycles
Pre-denaturation 95℃ 30 sec —
Denaturation 95℃ 10 sec 40 cycles
Annealing 55–65℃ 10 sec
Extension 72℃ 30 sec
Melting curve Instrument Default Instrument Default —

Recommended template loading amount is 1–2 μL; undiluted cDNA stock should not exceed 10% of the total reaction volume. Set annealing/extension temperatures based on primer Tm values; when the amplified fragment is ≤200 bp, the extension time can be set to 15 sec. Mix should be protected from strong light and gently mixed before use; do not vortex; no additional ROX is needed.

Assay Optimization and Primer Design

  • Primer Concentration
    Final concentration 0.1–1.0 μM. Lower concentrations increase specificity, but amplification efficiency slightly decreases; balance as needed.
  • Amplification Program
    To increase specificity, a two-step method or higher annealing temperature can be used; to increase amplification efficiency, a three-step method or longer extension time can be used.
  • Amplification Product
    Recommended length 80–200 bp.
  • Primer Parameters
    Primer length 18–23 bp; the Tm difference between the two primers ≤1℃, Tm controlled at 58–62℃; GC content 40%–60%.
  • Sequence Principles
    Try to keep the base distribution as even as possible, avoiding consecutive T/C or A/G structures; the 3' terminal preferably G or C, avoiding internal and primer-primer complementarity.
  • Specificity Verification
    Use NCBI BLAST search to confirm primer specificity.

Frequently Asked Questions

Does the qPCR reaction require additional ROX?

Not needed. Mix contains Universal calibration dye, suitable for verified compatible models. For specific models and parameter settings, please refer to the manual. When preparing the system, just add primers and template.

Amplification curve not smooth?

Mostly caused by too weak fluorescence signal, generated after system calibration. Ensure that the premixed dye has not degraded and use qPCR-specific consumables with better fluorescence signal collection.

Amplification curve broken or declining?

Often due to high template concentration or baseline endpoint value greater than Cq value. Try reducing the baseline endpoint (Cq value -4) and reanalyzing the data.

Sudden drop in amplification curve of individual wells?

Usually caused by bubbles in the reaction tube. Ensure the Mix is completely dissolved and do not vortex; gently tap and centrifuge after adding samples; pre-denaturation can be extended to 10 minutes.

No amplification curve at the end of the reaction?

Check whether the number of cycles has reached 40, whether the fluorescence collection step is correct, whether the primers have degraded, and whether the template concentration is too low or if the template has degraded.

Cq value appears too late?

This may be caused by low amplification efficiency, too low template concentration, template degradation, products being too long, or PCR inhibitors; you can increase primer concentration, switch to a three-step method, redesign primers, or increase the template dilution factor.

Multiple peaks in the melting curve?

It could be due to poor primer design or too high concentration, or genomic contamination in the cDNA template. You can redesign the primers, reduce the concentration, or use DNase to remove genomic contamination.

Poor experimental reproducibility?

Pay attention to pipetting errors, template concentration, and temperature differences between instrument wells. Use precise pipettes and high-quality tips, increase the reaction volume if necessary, and calibrate the instrument regularly.

Signal appears in the blank control?

It may be system contamination or primer dimers. First, replace the water used for the blank control. If the problem persists, replace primers, tips, PCR tubes, or use a new Mix, and prepare it in a clean bench.

Is the two-step method or the three-step method better?

The conventional recommendation is the two-step method; specificity can be increased by raising the annealing temperature or using the two-step method, and amplification efficiency can be improved by using the three-step method or extending the extension time.

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