StarLighter Dye-Based qPCR Master Mix (Universal)

• The amplification is highly specific and the data results are reliable;
• High amplification efficiency and early Ct value;
• High signal-to-noise ratio and high sensitivity;
• Fragments with high GC or high AT have good amplification effects;
• Strong tolerance to inhibitors.

• Gene expression analysis
• Low copy number gene testing;
• Gene knockout validation;
• Gene expression verification
• RNA copy number detection (virus and pathogen detection, etc.).

• Strong amplification specificity and excellent performance

Compared with many domestic and foreign brand qPCR Mix, Qihengxing SYBR Green qPCR Mix shows superior performance.

Fig.1 Comparison of qPCR results accuracy using different reagents

Using human cDNA as a template and reagents from the same manufacturer, a 219-bp fragment with a GC% of 64% (TR gene) was amplified. Amplification and melting curves were obtained for the StarLighter SYBR Green qPCR Mix and various brands. The SL-StarLighter SYBR Green qPCR Mix produced excellent amplification curves, strong fluorescence signals, and single, accurate melting curves and electrophoretic bands.

• High amplification efficiency and good linear range. High sensitivity for low copy detection

Fig.2 Comparison of quantitative results of different reagents for continuous gradient dilution template

Amplification curves and melting plots were obtained using the 8.3 pM standard from the KAPA Ion Torrent Library Quantification Kit as template (S1). Five-fold dilutions were then used to generate templates (S2-S10). qPCR quantification was performed using the StarLighter SYBR Green qPCR Mix and the supplier KP. The StarLighter SYBR Green qPCR Mix demonstrated good amplification efficiency (Eff = 100.483%) from S1-S10, with a perfect amplification curve, strong fluorescence signal, and a single melting curve.

• Strong fluorescence signal and high sensitivity

Fig.3 Comparison of fluorescence intensity of amplification curves of different reagents

Using the same template and primers, amplification curves were obtained using qPCR reagents from various brands on the market. The figure above compares the fluorescence signal values of various brands after removing ROX. The supplier KP and StarLighter SYBR Green qPCR Mix have the highest fluorescence signal intensity.

• Ct value appears earlier

Fig.4 Comparison of Ct values of qPCR results using different reagents

Using human DNA as a template and reagents from different manufacturers, a 219 bp fragment with a GC% of 64% (TR gene) was amplified. After electrophoresis confirmed the product as the target fragment, the StarLighter SYBR Green qPCR Mix achieved the lowest Ct value and the highest sensitivity after removing the Rox correction from the amplification curve.

• High GC or high AT fragments have good amplification effects

Fig.5 Comparison of the results of quantitative analysis of high AT fragments using different reagents

Using human DNA as a template and reagents from different manufacturers, a 198 bp fragment was amplified with an AT% of 64.4% (hPRT1 gene). The StarLighter SYBR Green qPCR Mix achieved the best amplification efficiency, with an Eff of 100.868%. Suppliers TY3 and KP achieved an Eff of 94.630% and 98.334%, respectively.

Fig.6 Comparison of quantitative results of high GC fragments using different reagents

Using human DNA as a template, reagents from different manufacturers were used to amplify a 322 bp fragment with a GC% of 75.5% (ApoE gene). The StarLighter SYBR Green qPCR Mix achieved the best amplification efficiency, with an Eff of 101.307%. Supplier TY3 achieved an Eff of 150.097%, while supplier AB achieved an Eff of NA (no amplification at the highest concentration).

• Strong tolerance to inhibitors

Fig.7 Comparison of quantitative results of inhibitor-containing templates using different reagents

Using human DNA as a template, reagents from various manufacturers were used to amplify a 393-bp fragment with a GC% of 35.6% (F8-exon 10 gene). DNA extracted using the Tiangen Magnetic Bead Blood Genomic Extraction Kit was used as template, with an initial template concentration of 100 ng/µL, as S1. Subsequently, S2, S3, and S4 were obtained by serial dilution using the same diluent. In terms of amplification efficiency, the StarLighter SYBR Green qPCR Mix achieved the best amplification efficiency of 94.365%, while the KP product achieved an Eff of NA (no amplification at the highest concentration). The StarLighter SYBR Green qPCR Mix achieved the required amplification efficiency, while the KP product's S1 template did not amplify.

• Reagents are stable and easy to store

Storage stability at 4°C

Fig 8 A

Fig 8 B and Fig 8 C

Fig 8 D
Fig.8 StarLighter SYBR Green qPCR Mix stability test results

Figure A: The reagent was stored at 4°C for up to 39 days; Figure B: The reagent was stored at room temperature for up to 8 days; Figure C: The reagent was stored at 31°C for up to 8 days; Figure D: The reagent was repeatedly frozen and thawed up to 35 times; using human DNA as the template, a 123 bp fragment (UBC gene) was amplified with a GC% of 52%, and no significant decrease in signal intensity or delay in C t value was observed.