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| Product Name | Advanced qPCR SYBR Master Mix (Universal ROX) |
| Catalog No. | RESK-0007 |
| Description | Advanced qPCR SYBR Master Mix (Universal ROX) is a 2× premix for real-time quantitative PCR featuring high fluorescence intensity, high sensitivity, strong specificity, and high amplification yield. The mix is blue in color, providing a built-in pipetting tracking aid during reaction setup. Its core component, an antibody-based hot-start Taq DNA polymerase, effectively suppresses nonspecific amplification caused by primer annealing during sample preparation. The formulation also contains factors that boost amplification efficiency and balance the amplification of genes with different GC contents (30%-70%), enabling excellent linearity across a broad quantification range. |
| Intended Use | Dye-based real-time quantitative PCR for sensitive and specific quantification of DNA/cDNA templates, including low-copy-number targets, across a wide range of qPCR instruments. For research use. |
| Principle / Technology | SYBR Green dye-based fluorescent detection combined with an antibody-based hot-start Taq DNA polymerase: the blocking antibody keeps the polymerase inactive at low temperatures, preventing nonspecific amplification from primer annealing during reaction setup, and is released upon heat activation. An optimized buffer further minimizes primer-dimer formation, while enhancer factors improve amplification efficiency and equalize yields across templates with 30%-70% GC content. A universal ROX formulation allows use on most qPCR platforms without adjusting ROX concentration. |
| Detection Method | Fluorescent quantitative PCR (SYBR Green dye-based) |
| Sample Type | DNA or cDNA templates (e.g., plasmid DNA, human cDNA) with gene-specific primers; recommended amplicon length 80-300 bp; applicable to templates of varying GC content (30%-70%). |
| Performance Range / Specifications | Reliable amplification across a wide dynamic range, validated with 10^1-10^7 copies of plasmid template on different instrument types. Accurately discriminates 2-fold differences in template concentration (a 5-fold concentration difference corresponds to a ΔCt of 2.322). Low nonspecific amplification rate: approximately 5.4% across 56 human gene fragments of varying GC content, versus approximately 53.6% for other brands. High plateau fluorescence, better curve shape, and higher signal-to-noise ratio; tight agreement between replicates. Compatible with most qPCR instruments without ROX concentration adjustment. |
| Sensitivity / LOD | Effectively detects low-copy-number templates, validated down to 10^1 copies of plasmid DNA, with lower Ct values and higher fluorescence plateaus than competing products. |
| Specificity | Antibody-based hot start combined with buffer optimization minimizes primer-dimer formation; nonspecific amplification probability is approximately 5.4% across tested human gene panels, far below competing products (approximately 53.6%). |
| Reaction Conditions / Protocol | Standard qPCR cycling per instrument requirements; validated across multiple qPCR platforms without ROX adjustment. For viscous mix handling, prepare a bulk master mix, dispense into tubes, and add template last; dispense slowly along the tube wall and avoid introducing air bubbles (spin tubes or tap plates gently to remove residual bubbles). |
| Components / Formulation | 2× Advanced qPCR SYBR Master Mix (Universal ROX), blue-colored, containing antibody hot-start Taq DNA polymerase, SYBR dye, dNTPs, Mg2+, universal ROX reference dye, and stabilizers Package sizes: 1 mL; 5× 1 mL; 100× 1 mL |
| Storage Conditions | Store at -25 to -15 °C protected from light. A pale-yellow precipitate may form after frozen storage; this is a stabilizer with reduced solubility at low temperature and does not affect performance. After thawing, invert and pipette to mix until the precipitate disappears; if it persists, centrifuge and use the supernatant without taking up the precipitate. |
| Shelf Life | 18 months when stored at -25 to -15 °C protected from light. |
| Package Specifications | 1 mL 5× 1 mL 100× 1 mL |
| Product Form | 2× ready-to-use liquid premix (blue, with pipetting tracking dye) |
| Concentration | 2× |
| Source / Origin | Antibody hot-start Taq DNA polymerase with optimized buffer system, SYBR dye, dNTPs, and universal ROX reference dye. |
| Expiration Date / Stability | Stable for 18 months at -25 to -15 °C protected from light. |
| Compatibility | Suitable for the vast majority of fluorescence quantitative PCR instruments; universal ROX formulation requires no ROX concentration adjustment across platforms. |
| Application Notes / Precautions | Smearing on post-amplification gel electrophoresis is caused by inert stabilizer components in the reagent that do not participate in the reaction; running qPCR products on a gel after cycling is not recommended. The mix is viscous and prone to bubbles: prepare a bulk mix first, then aliquot into tubes and add template last; aspirate at second stop and dispense at first stop, expel liquid slowly without blow-out, and dispense along the tube wall. Remove residual bubbles by brief centrifugation of PCR tubes or by gently tapping 96-well plates. A pale-yellow precipitate after removal from the freezer is precipitated stabilizer and does not affect performance; mix by inversion and gentle pipetting until dissolved, or centrifuge and avoid drawing the precipitate. Compared with standard formulations, this advanced mix offers better specificity, higher sensitivity, and higher plateau fluorescence signal. |
For research use only, not for clinical use.
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