Transcriptor One-Step RT-PCR Kit

Description

• General description: For hot start one-step RT-PCR, utilizing the enzyme mixture of Transcriptor Reverse Transcriptase, Protector RNase Inhibitor, and the Expand System, with an innovative hot start buffer (patents pending).
• Application: The Transcriptor One-Step RT-PCR Kit is designed for fast, sensitive, and specific end-point RT-PCR analysis of RNA (total RNA, mRNA, in vitro-transcribed RNA, or viral RNA) using gene-specific primers.Featuring an innovative reaction buffer, the kit combines sensitive, high-yield reverse transcription with the improved performance of a hot start system for high-fidelity, high-yield amplification of a variety of templates, including GC-rich RNA.The kit offers a convenient, highly sensitive one-step RT-PCR method to generate transcripts up to 6.5 kb for applications such as: Qualitative, semi-quantitative, or quantitative analysis of RNA transcription levelsDetermining the presence or absence of RNA templatesQuantifying gene expression through RNA analysis (e.g., for virus-level quantification)Cloning of RNA up to 6.5 kbThe one-step format is convenient, reduces reaction-to-reaction variability, and prevents contamination by minimizing hands-on steps.
• Features and Benefits: Obtain high sensitivity and yield in a wide amplification range:Detect as little as 1 fg of total RNA.Achieve high specificity and reduce primer-dimersCount on excellent performance with the kit′s proprietary hot start buffer (patents pending).Produce long fragments:Generate transcripts up to 6.5 kb.Transcribe a variety of templates, even the most difficult (e.g., GC-rich RNA):Overcome high secondary structure with thermostableTranscriptor Reverse Transcriptase in combination with an optimized buffer.Save time and improve results with a convenient kit:Minimize pipetting steps and prevent contamination by using a premixed enzyme blend and optimized buffer that includes PCR-grade dNTPs and hot start components.Protect your RNA sample:Safeguard your RNA from degradation by RNases with the supplied thermostable Protector RNase Inhibitor that is functional during one-step RT-PCR when other RNase inhibitors fail.Increase confidence in your results:Use the supplied control reactions to monitor performance.
• Components: RT-PCR Enzyme Mix containing Transcriptor Reverse Transcriptase, Protector RNase Inhibitor, and the Expand SystemRT-PCR Reaction Buffer containing hot start mediating components and dNTPs – 5x concentratedWater, PCR GradeControl RNA – HAV RNA, in vitro transcript *Control primer mix HAV RNA (forward and reverse primer) ** Included only in the 50-reaction pack size.
• Quality: Each lot of the Transcriptor One-Step RT-PCR Kit is function tested in RT-PCR. RT-PCR is performed with 1000 copies of HAV RNA, in vitro transcript (also provided with Cat.No 04 655 877 001). The control reaction setup and the subsequent RT-PCR with primers for a 246 bp HAV RNA fragment are performed using the standard RT-PCR protocol (reverse transcription at +50°C for 30 minutes) provided in the package insert. With 1,000 copies of target a clearly visible band is obtained after agarose gel electrophoresis and ethidium bromide staining.In addition, RT-PCR is performed on a dilution series of human liver total RNA (10 ng, 1 ng, and 0.1 ng). RT-PCR with specific primers for a 2.5 kb fragment of ApoB is performed using the standard RT-PCR protocol (reverse transcription at +50°C for 30 minutes) provided in the package insert. With 1 ng of RNA a clearly visible band is obtained after agarose gel electrophoresis and ethidium bromide staining.
• Other Notes: Reverse Transcriptases Overview