SOLIScript® Fast 1-step RT-qPCR Mix with UNG
4Pages

SOLIS BIODYNE OÜ A: Teaduspargi 9, 50411 Tartu, Estonia E: info@solisbiodyne.com T: +372 7409 960 F: +372 7402 079 SOLIScript Fast 1-step RT-qPCR Mix with UNG ® SOLIScript® Fast 1-step RT-qPCR Mix with UNG is optimized for probe-based one-step RT-qPCR assays. It contains all components necessary in one tube (except template and primers) to perform cDNA synthesis and qPCR with up to 5-targets within around 1h of total reaction time. It also contains the RNase Inhibitor RiboGrip™ to protect RNA sample from degradation. Inhibitor tolerance and fast cycling allow flexible experiment design. Salini UNG™ Uracil-N-Glycosylase is incorporated into the product along with dUTPs to prevent carryover contamination and false positive results. RT-qPCR reactions with two products, SOLIScript® Fast 1-step RT-qPCR Mix with UNG and a version without UNG were spiked with equal concentration of dU-containing amplicons, mimicking carryover contamination. While the reagent without UNG generated a regular amplification curve (purple, Figure 2), SOLIScript® Fast 1-step RT-qPCR Mix with UNG degraded the dU-containing amplicons (red, Figure 2), resulting in no amplification from the mimicking carryover contamination. Routine storage at -20°C Shipping conditions: at room temperature Prevent carryover contamination Inhibitor tolerance HOT SolisFAST® DNA Polymerase is an in silico designed analogue of Taq DNA polymerase with enhanced stability at room temperature, chemical hot-start, increased tolerance to inhibitory substances and approximately 2-4 times faster extension rates compared to the wild-type Taq DNA polymerase. SOLIScript® RT is an in silico-engineered thermostable reverse transcriptase that retains activity at higher temperatures (up to 60°C) to provide specific results when working with templates with high level of secondary structures. Wide dynamic range Fast cycling SOLIScript® Fast 1-step RT-qPCR Mix with UNG Figure 1. PPIA target from human total RNA was amplified over eight 10-fold dilutions (1000 ng to 100 fg, R2=1.0), showing sensitive detection over a wide dynamic range. Reactions were ran on Bio-Rad CFX96 platform. Figure 2. Amplification plot showcasing results with SOLIScript® Fast 1-step RT-qPCR Mix with UNG (red) and a formulation without UNG (purple). Reactions were ran on Bio-Rad CFX

Effective 5-plex amplification Five-plex RT-qPCR reactions with SOLIScript® Fast 1-step RT-qPCR Mix with UNG show strong amplification and consistent efficiencies across all five targets with different charateristics. Amplification Figure 3. Five-plex RT-qPCR reactions (FAM, blue HEX, green; ROX, red; Cy5, puprle; Cy5.5, orange) on Bio-Rad CFX96 platform with six 10-fold serial dilutions of reference human total RNA (from 1000 ng/µl to 0.01 ng/µl). Simultaneous amplification of RNA and DNA targets 40 A test system for simultaneous detection of RNA and DNA targets was created. Four 10-fold...

Solis BioDyne Figure 5. Five-plex RT-qPCR reactions were performed with SOLIScript® Fast 1-step RT-qPCR Mix with UNG and three competitor products. Six 10-fold dilutions of human total RNA (from 100 ng/rxn to 1 pg/rxn) were tested. Reactions were ran on the Bio-Rad CFX96 platform and reaction conditions were chosen according to each manufacturer’s instructions. (A) Cq values from the 5-plex reactions (results from FAM and HEX channel presented). (B) Amplification plots from the 5-plex reactions (results from HEX channel presented). Five-plex RT-qPCR reactions with SOLIScript® Fast 1-step...

Lot-to-lot consistency Figure 6. Results from lot-to-lot consistency tests with 4-plex RT-qPCR reactions over three 10-fold RNA dilutions (0.1 ng/rxn, orange; 0.01 ng/rxn, green; 0.001 ng/rxn, blue). Mean Cq values across three different production lots along with standard deviations are presented (if SD>0.16). Mean Cq values of three lots Results from lot-to-lot consistency tests showcase that production under strict quality systems ensures reliable consistency between independent production lots of SOLIScript® Fast 1-step RT-qPCR Mix with UNG. Solis BioDyne About Solis BioDyne Established...