HelixCript™ One-Step RT-PCR Kit [Hot-Taq] is designed for highly specific amplification of target genes in one-tube reactions from total transcripts. A reverse transcriptase, an RNase Inhibitor, and Hot-Taq polymerase (chemically-modified) are the supplied enzyme mixture. The 2x Reaction Mix contains a unique buffer, MgCl₂, and dNTPs.
Figure 1. Multiplex one-step RT-PCR using HelixCript™ One-Step RT-PCR Kit [Hot-Taq]. Multiplex one-step RT-PCR in a single-tube reaction was performed with total RNA extracted from a hot pepper plant (Capsicum annuum) and then adding the indicated gene-specific primer sets. Each lane from 1 to 4 represents the progressive number of primer sets. The targets of the used primer sets are; CaAPX1 (374 bp), CaActin (461 bp), CaPF1 (615 bp), and Cadhn (761 bp).
Figure 2. Viral RNA detection using HelixCript™ One-Step RT-PCR Kit [Hot-Taq]. Total RNA was isolated from a nasopharyngeal specimen of a new influenza virus-infected patient. Each reaction was performed with one of these target-specific primer sets: influenza A (InfA, 106 bp), Swine influenza A (SW InfA, 195 bp), Swine Influenza H1 (SW H1,116 bp), or RNase P (human internal control, 65 bp).
Application
- Detection of target gene transcript from RNA
- Cloning of target cDNA products
- Semi-quantitative, quantitative analysis of RNA transcription level
Contents
- Enzyme Mix [Hot-Taq], 2x Reaction Mix [Hot-Taq]
Quality Control
The activity for cDNA synthesis and PCR amplification of target gene transcript using the HelixCript™ One-Step RT-PCR Kit [Hot-Taq] was evaluated by Limit Of Detection (LOD) assay from 10-fold serial dilutions of human total RNA.
