QIAGEN OneStep - Model RT-PCR Kit -Sensitive And Successful One-Step RT-PCR For Viral RNA Detection

For highly sensitive and successful one-step RT-PCR

• Fast and easy one-tube setup
• One-step RT-PCR of any RNA template without optimization
• Unique enzyme mix for high specificity and sensitivity
• Optimized reverse-transcription and amplification buffer

The QIAGEN OneStep RT-PCR Kit is designed for easy and sensitive one-step RT-PCR using any RNA template. A unique enzyme combination and specially developed reaction buffer ensure efficient, highly specific reverse transcription and PCR in one tube, without the need for optimization.

The QIAGEN OneStep RT-PCR Enzyme Mix contains a specially formulated enzyme blend for both reverse transcription and PCR. The unique combination of Omniscript and Sensiscript Reverse Transcriptases, with their high affinity for RNA templates, ensures highly efficient and sensitive transcription of RNA amounts from as little as 1 pg up to 2 µg. After reverse transcription, reactions are heated to 95°C for 15 minutes to activate HotStarTaq DNA Polymerase and simultaneously inactivate the reverse transcriptases. This hot start to the PCR eliminates any nonspecific amplification products such as primer-dimers and reduces background smear, ensuring highly sensitive and reproducible RT-PCR.

The optimal primer annealing temperature is dependent on the base composition (i.e., the proportion of A, T, G, and C nucleotides), primer concentration, and ionic reaction environment. Using QIAGEN PCR Buffers, containing both K+ and NH4+, delivers high yields of specific PCR product over a wide range of annealing temperatures. This specificity is achieved by destabilizing non-specifically bound primers, providing a more robust reaction environment and eliminating the need for tedious annealing temperature optimization. In contrast, the range of optimal PCR annealing temperatures is smaller and less predictable using a PCR or one-step RT-PCR buffer that only contains K+, as illustrated in figure "Influence of Annealing Temperature on One-Step RT-PCR Specificity". View this animation and discover the benefits of QIAGEN`s unique dual-cation PCR buffer.

QIAGEN OneStep RT-PCR Buffer has been specially developed to allow both efficient reverse transcription and PCR amplification. The buffer contains novel additives that prevent inhibition of PCR amplification by reverse transcriptases, a problem often encountered in one-step RT-PCR. The buffer ensures specific primer annealing over a wide range of temperatures and Mg2+ concentrations; providing robust and highly efficient RT-PCR from any RNA template. The QIAGEN OneStep RT-PCR Kit includes Q-Solution, an innovative additive that modifies the melting behavior of nucleic acids. Q-Solution facilitates reverse transcription and amplification of templates with a high GC content or a high degree of secondary structure. Using Q-Solution simplifies optimization of RT-PCR for difficult templates. The QIAGEN OneStep RT-PCR Kit comes with everything you need for faster and easier RT-PCR for your most sensitive applications.

The QIAGEN OneStep RT-PCR Kit allows fast and easy RT-PCR setup. Whatever your application — virus detection, molecular diagnostics research, or gene expression — just mix all components together in one tube and start your thermal-cycler program . The reaction mixture contains all of the reagents required for both reverse transcription and PCR; nothing needs to be added once the reaction has been started.

The QIAGEN OneStep RT-PCR Kit is suitable for RT-PCR applications such as:

• Virus detection
• Single-cell RT-PCR
• Gene-expression analysis