The Most Reliable SYBR® Green-based Real-Time qPCR Assays for Gene Expression Analysis

RT² qPCR Primer Assays are the mos...
Why Use RT² qPCR Primer Assays?

RT² qPCR Primer Assay Features

SYBR® Green assays do not require the design of a specifi...
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Rt2q pcr primerassays

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Rt2q pcr primerassays

  1. 1. The Most Reliable SYBR® Green-based Real-Time qPCR Assays for Gene Expression Analysis RT² qPCR Primer Assays are the most reliable gene expression analysis tool using SYBR Green-based quantitative real-time PCR Verified Specificity Ct = 27.3 Ct = 21.1 technology. Each assay utilizes a proprietary and experimentally verified algorithm for designing gene-specific qPCR primers with uniform PCR efficiency and amplification conditions. Every RT² qPCR Primer Assay is subjected to rigorous experimental verification. Amplification of a single product of the correct size and high PCR efficiency (>90%) are guaranteed when the assays are used with the RT² qPCR Master Mixes. With high PCR efficiency and uniform PCR conditions, it is convenient to perform quantitative real-time PCR for several genes under the MMP13 MMP15 Each RT² qPCR Primer Assay is experimentally verified to yield a single dissociation curve peak and to generate a single amplicon of the correct size for the targeted gene. Wide Linear Dynamic Range same PCR conditions. Benefits of RT2 qPCR Primer Assays High Performance: Each RT² qPCR Primer Assay is experimentally validated to amplify a single amplicon of correct size at uniform PCR efficiency. The performance of RT² qPCR Primer Assays is similar to that of TaqMan® assays. Complete Genome Coverage: RT² qPCR Primer Assays are available for every human, mouse and rat gene. Their uniform PCR efficiency and PCR conditions allows easy transition from single gene analysis to multiple gene expression profiling. Convenience: With less than 5-minutes of hands-on time, RT² qPCR Primer Assays deliver guaranteed performance when used with optimized RT² qPCR Master Mixes. Master Mixes are available for all real-time PCR instruments. Wide linear dynamic range enables simultaneous analyses for rare and abundant transcripts. Ten-fold serial dilutions of purified DNA were used as template in RT² qPCR Primer Assays. The qPCR was run in duplicate for each dilution point. The RT² qPCR Primer Assays detect from 1 copy to 1010 copies of template covering a linear dynamic range of up to eight (8) orders of magnitude. Equivalent Performance to TaqMan Assays TaqMan Gene Expression Assays Fold Change (log2) qPCR ASSAYS RT2 qPCR Primer Assays 20 15 10 5 -20 -15 -10 -5 5 10 15 20 -5 -10 -15 -20 RT2 qPCR Primer Assays Fold Change (log2) A high concordance is observed between RT² qPCR Primer Assays and TaqMan Gene Expression Assays. Gene expression comparisons were made between two samples for 84 genes by the RT² qPCR Primer Assay method and the TaqMan Gene Expression Assay method. Fold changes (log 2 ) in expression for each gene are calculated and plotted against one other for the two methods. The gray dashed line represents a straight line with an ideal slope of 1.0. The solid black line shows the linear regression data fit. The Pearson correlation coefficient (R value ) between the two sets of data is 0.97.
  2. 2. Why Use RT² qPCR Primer Assays? RT² qPCR Primer Assay Features SYBR® Green assays do not require the design of a specific probe High Specificity Single band of correct size in addition to the primers for each gene-specific assay, reducing Uniform Efficiency >90% PCR efficiency* the assay setup time and running costs. A drawback of SYBR High Sensitivity As low as 10 copies per cell Green assays, however, is that the dye binds all double-stranded Wide Dynamic Linear Range 10 - 109 copies DNA non-specifically and can generate false positive signals if non-specific products and primer dimers are present in the assay. Conventional PCR primers, not specifically designed for Applications real-time PCR, often have the problems of low PCR amplification efficiency, primer dimers, and non-specific amplification. In Validate microarray-derived gene expression data. short, conventional PCR primers can not be used for real-time PCR. Examine a focused panel of genes using multiple RT² qPCR Primer Assays. RT² qPCR Primer Assays are specifically designed for real-time PCR analysis. The assay design criteria ensure that each qPCR Verify gene expression knock down by RNAi. reaction will generate single, gene-specific amplicons and prevent the co-amplification of non-specific products. Furthermore, Identify and confirm disease-associated simultaneous gene expression analyses require similar PCR biomarkers. efficiencies for accurate comparisons among genes. RT² qPCR Monitor phenotypic changes related to gene Primer Assays are designed with an amplicon size range of 100- expression. 250 bp and their PCR efficiency* is uniformly >90%. Overall, more than 10 thermodynamic criteria are included in each RT² qPCR Primer Assay design to deliver the most reliable and * PCR Efficiency is calculated using DART-PCR workbook (Nucleic Acid Research 2003 31 (14): e73.) accurate gene expression analysis results. RT2 qPCR Primer Assays Ordering Guide www.SABiosciences.com/RT2PCR.php Find the qPCR Assay for your genes at: Description Gene-Specific RT2 qPCR Primer Assay Cat. No. PPX#####Y-200 X = H, Human; M, Mouse; R, Rat #####Y identifies specific gene and design For Guaranteed qPCR Performance, Use RT2 Reagents: RT2 qPCR Master Mixes Instrument Type Cat. No. RT2 SYBR Green / ROX qPCR Master Mix Applied Biosystems, Stratagene, Roche PA-112 (2000 qPCR reactions) 2 RT SYBR Green / Fluorescein qPCR Master Mix Bio-Rad PA-111 (2000 qPCR reactions) RT2 SYBR Green qPCR Master Mix PA-110 (2000 qPCR reactions) RT2 First Strand Kit C-03 RT2 qPCR-Grade RNA Isolation Kit PA-001 (12 RNA isolations) All other real-time instruments (12 first strand reactions) RT2 qPCR Primer Assays™ and PCRGrade™ are trademarks of SABiosciences Corporation. SYBR® is a registered trademark of Molecular Probes, Inc. ABI® and ROX® are a registered trademarks of Applera Corporation. Bio-Rad® is a registered trademark of Bio-Rad Laboratories. Stratagene® is a registered trademarks of Stratagene. TaqMan® is a registered trademark of Roche Molecular Systems. SABiosciences Corporation 6951 Executive Way Frederick, MD 21703 www.SABiosciences.com T 301.682.9200 888.503.3187 F 301.682.7300 888.465.9859

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