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YOUR PA RT NER F OR... HIGH-PERFORM A NCE GENE E XPRESSION RNA Purification cDNA Synthesis Real-Time PCR I S O L AT E I I RNA KITS S ensi FA S T c D N A SYNTHESIS KIT S ensi FA S T P R O B E AND SYBR® KITS FROM SAMPLE TO RE AL-TIME RESULTS WITH BIOLINE BIOLINE PROVIDES HIGH-PERFORMANCE PRODUCTS FOR ALL OF YOUR GENE EXPRESSION STUDIES. Start with ISOLATE II Kits for purification and concentration of high-integrity total RNA from a wide range of starting materials. Continue with the SensiFAST cDNA Synthesis Kit to generate real-time ready cDNA for complete unbiased 5’ to 3’ RNA sequence representation. When used in combination with SensiFAST Probe and SYBR® Kits, which use the latest advances in buffer chemistry, Bioline delivers ultra-sensitive real-time results in less than an hour. ISOLATE II RNA KITS RNA Purification TOTAL RNA PURIFICATION RNA CLEAN-UP AND CONCENTRATION RNA MINI KIT Fast, efficient isolation of pure total RNA from a variety of sources. R N A M I C RO C L E A N - U P K I T Purify and concentrate RNA, free from enzymatic inhibitors or impurities. Innovative column design allows as little as 5μl elution volumes. RNA PL ANT KIT Pure total RNA isolation from plant tissues, including leaves, bark, roots and fruits. R N A M I C RO K I T For isolation of pure total RNA from small samples, including single cells and biopsy material. M1 • Simultaneous RNA clean-up and concentration • Concentrates RNA up to 50-fold. • Ideal for RNA purification following enzymatic digests or phenol-extracts. 2 34 5 67 M Fig. 1 High-quality RNA isolated from plant tissue RNA was isolated from 20mg freeze-dried budding leaves of Arabidopsis thaliana using ISOLATE II RNA Plant Kit. The extracted RNA was diluted in a 2-fold serial dilution (15ng, lanes 1-7 respectively) and PCR was performed using MyTaq One-Step RT-PCR Kit. HyperLadder™ 1kb (M). The results illustrate the quality of the RNA obtained, as it can be used for very sensitive cDNA synthesis and PCR without further purification. Fig. 2 Consistent RNA clean-up RNA was isolated from HeLa cells using standard phenol extraction split into eight 22μg samples and purified using the ISOLATE II RNA Micro Clean-Up Kit. The cleaned RNA samples were run on an Agilent Bioanalyzer 2100. All RIN values were >9, indicating highly pure RNA and ideal for downstream applications. SensiFAST™ cDNA SYNTHESIS KIT cDNA Synthesis GENERATE REAL-TIME READY FIRST STRAND cDNA FROM TOTAL RNA TEMPLATE. • Reproducible - Unique reverse transcriptase and new TransAmp buffer generate consistent first-strand cDNA • Broad dynamic range - ideal for dilute and low-copy samples • Sensitive - real-time RT-PCR analysis from as little as 1pg of starting total RNA • Unbiased cDNA synthesis - complete 5’ to 3’ RNA sequence representation • Rapid - 30 minute cDNA Synthesis protocol, ideal for high throughput applications ∆Rn Amplification Plot Cycle Fig. 3 SensiFAST cDNA Synthesis kit gives unbiased synthesis Primer pairs designed at 1kb intervals across the same transcript. Unlike Supplier B (blue), the SensiFAST cDNA Synthesis Kit (red) has no bias across a 5kb transcript. Amplification curves Fluorescence (465-510) The SensiFAST cDNA Synthesis Kit has been developed to generate real-time ready first strand cDNA from total RNA template. The kit contains all necessary components in the unique TransAmp™ buffer system to deliver the highest quality cDNA. An optimized blend of random hexamer and anchored oligo(dT) primers provide robust, consistent and unbiased first-strand synthesis over a broad range of RNA template concentrations, making SensiFAST cDNA Synthesis Kit the ideal partner for SensiFAST Probe and SYBR® realtime PCR kits. 100ng 100pg Cycles Fig. 4 Reproducibility of SensiFAST cDNA Synthesis Kit SensiFAST cDNA Synthesis Kit was employed in 48 independent first-strand reactions in triplicate, containing 100ng or 100pg of total RNA. The results demonstrate the excellent reproducibility of SensiFAST cDNA Synthesis Kit (the same Ct values), across the 144 wells with high levels and the 144 wells with low levels of input target mRNA. SensiFAST™ PROBE AND SYBR ® KITS Real-Time PCR REAL-TIME RESULTS IN LESS THAN AN HOUR. SensiFAST uses the latest developments in real-time PCR to realize the fastest cycling times and greatest sensitivity, without compromising accuracy, reproducibility or performance. • Fast - optimized for fast cycling conditions (under 30 minutes) • High specificity - leading to better efficiency and more confidence in your results • Ultra-sensitive - perfect for low-copy number samples • Universal - compatible with all standard and fast cycling instruments. A. B. Fig. 7 Superior performance in multiplex assays A 10-fold serial dilution of human cDNA was amplified with 4 different probes (results displayed for a single gene), both in singleplex reactions (blue line) and quadruplex reactions (red line). The results illustrate that SensiFAST Probe No-ROX has high sensitivity and excellent reproducibility for both singleplex and multiplex reactions (Fig. 7A) and no reduction of efficiency (Fig. 7B) that is often associated with multiplexing. There is no change in Ct values between singleplex and multiplex reactions. Fig. 5 Superior speed and specificity under fast cycling conditions A 10-fold serial dilution of human genomic DNA was amplified using SensiFAST Probe Lo-ROX (red line) and the results were compared with amplifications using a kit from Supplier B (blue line) under fast cycling conditions (45 cycles of 95°C for 10s, 60°C for 10s). The results illustrate the quality of SensiFAST Probe Lo-ROX, which is faster (earlier Ct) and more sensitive than Supplier B. Fig. 6 High sensitivity, enabling detection of expressed genes 10-fold serial dilutions of human genomic DNA (in triplicate) over 5 orders of magnitude was amplified using SensiFAST Probe No-ROX (red line) and the results were compared with amplifications using a kit from Supplier R (blue line). Results illustrate that SensiFAST Probe is at least 10 fold more sensitive in this assay. “ ” HERE IS WHAT OTHER RESE ARCHERS SAY ABOUT OUR PRODUCTS “SensiFAST ™ consistently exhibits excellent sensitivity in our differential gene expression assays and superior efficiency compared with our previous qPCR mix.” National Institutes of Health Bethesda, Maryland, USA FOR MORE INFORMATION PLEASE VISIT bioline.com/genex P G G B L 0714V1.1