Features & Benefits

• Supports multiple priming strategies - oligo (dT), random primers, or gene-specific primers
• Proprietary enhancer solution improves priming efficiency for higher cDNA yield
• Broad dynamic range
• Engineered MMLV reverse transcriptase plus RNase inhibitor prevents post-synthesis degradation of RNA template
• High quality, high efficiency cDNA synthesis, up to 8 Kb+

Description

The qScript™ Flex cDNA Synthesis Kit is an easy-to-use and highly efficient kit for the synthesis of first-strand cDNA that enables your choice of cDNA priming method. The kit provides optimized reagents for priming with oligo(dT)20, random primer, gene-specific primer (GSP), or any combination thereof. A key component is a proprietary enhancer compound that improves cDNA priming efficiency. This is provided as a separate solution for use with your gene-specific primers, or pre-blended with the supplied Oligo dT or Random Primer solutions. qScript™ reverse transcriptase is a mixture of an engineered RNase H+ MMLV RT and recombinant ribonuclease inhibitor protein that has been specifically optimized for use with the provided qScript™ Flex Reaction Mix. This unique 5X master mix of buffer, magnesium, stabilizers and dNTPs simplifies reaction assembly and ensures robust and reproducible synthesis of first strand product from 10 pg to 1µg of total RNA or purified polyA+ RNA template. The resulting cDNA product is directly compatible with current real-time RT-PCR methods or end-point RT-PCR. Length of cDNA product is dependent upon priming strategy and quality of the RNA template. Oligo dT or GSP can be used for long RT-PCR of RNA targets up to 12 kb. Random primer is suitable for RT-PCR of RNA targets less than 1 kb. Additionally, the location of the target sequence in the mRNA should be considered when selecting a priming method. Oligo dT primer is the preferred choice for RT-PCR of 3' end sequences. The efficacy of RT PCR of 5' end regions is often improved by using random or gene-specific primers. Any of the three priming methods is suitable for real-time quantitative RT PCR (qRT-PCR). However, we recommend using a mixture of the Oligo dT and Random Primer to achieve consistent and unbiased first-strand synthesis over a broad range of RNA template concentrations. This priming strategy has been incorporated into our qScript™ cDNA SuperMix (95048-025, 95048-100, 95048-500) and qScript™ cDNA Synthesis Kit (95047-025, 95047-100, 95047-500). These kits provide streamlined optimized solutions for gene-expression studies using real-time RT-PCR.

Contents

• qScript Flex Reaction Mix (5X) - 5X concentrated solution of optimized buffer, magnesium, dNTPs, and stabilizers
• Oligo dT - 10X concentrated solution of oligo(dT)20 with GSP Enhancer
• Random Primer - 10X concentrated solution of random primer with GSP Enhancer
• GSP Enhancer (10X)
• qScript Reverse Transcriptase
• Nuclease-free water

Storage conditions

Kit components are stable for one year when stored in a constant temperature freezer at -20°C. After thawing, mix thoroughly before using.