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Thermo Scientific Solaris qPCR Master Mix was developed in conjunction with Solaris qPCR Gene Expression Assays for quantification of DNA and cDNA. Optimal qPCR results using probe detection chemistries can be achieved through this system of qPCR products. These optimized master mixes offer a unique feature – they’re blue. Working with blue reagents allows you to see your reagent as you pipette, track your progress across multiple wells, and visually assess your set-up. This visual confirmation further enhances the repeatability of your data.
The Solaris qPCR Master Mix, in combination with Solaris Gene Expression Assays, enables analysis of gene expression in human and mouse.
With the exception of probe, primers and template the Solaris qPCR Gene Expression Master Mix contains all the components required for reliable, sensitive and specific qPCR detection. These components include:
Thermo-Start DNA Polymerase requires a 15 minute activation step at 95°C.
For maximum sample purity, total RNA should be extracted using a spin column method. The Thermo Scientific Verso cDNA Synthesis Kit should be used in the upstream reverse transcription (RT) step for improved cDNA yield.
Solaris qPCR Master Mix and Solaris qPCR Gene Expression Assays are compatible with qPCR platforms from all major suppliers, including;
Solaris gives highly reproducible assay results, as judged by the PCR efficiency and r2 values. PPIB was amplified in quadruplicate from six ten-fold dilutions of cDNA synthesized from 100 ng total RNA on three instruments, ABI 7900HT (384-well format), Roche LightCycler 480 (384-well format) and Stratagene Mx3000P (96-well format). Calculated efficiency and r2 values are shown on each amplification curve. The average Cq value for the highest amount of input cDNA for the three instruments is 20.5 with standard deviation of ± 0.6.
Solaris Assays give reliable detection even at very low input concentrations, as judged by the PCR efficiency and r2 values. Ten 10-fold dilutions of cDNA synthesized from synRNA amplicon sequence or DNA amplicon sequence was amplified on an ABI 7900HT instrument using Solaris qPCR Gene Expression Assay for F2RL1 or CDC20, respectively. The log-scale amplifi cation curves and standard curves are shown along with the performance of each assay including efficiency, r2 value, dynamic range out of 10 log10 dilutions and the lower limit of detection.
Solaris gives efficient, repeatable detection of all gene targets on all commonly used qPCR platforms and targets.
Solaris Assays give consistent results even between different researchers and laboratories. siRNAs targeting ALDOA and PPIB, as well as a Non-targeting Control (NTC), were transfected into HeLa cells at 100, 10, 1 and 0.1 nM final concentrations. Cells were harvested and total RNA isolated 48 hrs post-treatment. cDNA was synthesized using Thermo Scientific Verso cDNA Synthesis Kit. An aliquot of the cDNA was amplified in two geographically separated laboratories using Solaris qPCR Gene Expression Assays for detection of ALDOA, PPIB, and GAPDH on a Roche LightCycler 480 (384-well) platform. Knockdown was calculated using the δδCq method (normalized to GAPDH reference gene and NTC-treated cells). The same levels of knockdown were demonstrated for both gene targets between both researchers at both sites.