HelixCript™ One-Step RT-PCR [Speed-Pfu] is designed for sensitivity amplification of target genes in one-tube reactions from total transcripts. HelixCript™ Thermo Reverse Transcriptase, Helixzyme™ RNase Inhibitor, and HelixAmp™ Speed-Pfu polymerase (fast PCR enzyme with high fidelity) are supplied as the enzyme mixture. The 2x Reaction Mix contains a unique buffer, MgCl₂, and dNTPs.

Figure 1. A fast RT-PCR using HelixCript™ One-Step RT-PCR Kit [Speed-Pfu]. One-Step RT-PCR reactions were performed under the modified protocol as follows : The total time required for a One-Step RT-PCR reaction is 1 hour. One-step RT-PCRs were perfomed with 1 to 100 ng of human blood total RNA using HelixCript™ One-Step RT-PCR Kit [Speed-Pfu] and the primer set for human β-globin. The PCR reaction was carried out at 95℃(10 sec), 55℃(10 sec), and 72℃(20 sec) for 35 cycles.

Application 

• Detection of target gene transcript from RNA 
• Cloning of target cDNA products
• Semi-quantitative, quantitative analysis of RNA transcription level 

Contents 

• Enzyme Mix [Speed-Pfu], 2x Reaction Mix [Speed-Pfu] 

Quality Control

The activity for cDNA synthesis and PCR amplification of target gene transcript using the HelixCript™ One-Step RT-PCR Kit [Speed-Pfu] was evaluated by Limit Of Detection (LOD) assay from 10-fold serial dilutions of human total RNA.