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HelixCript™ One-Step RT-PCR Kit
● Single-tube reaction
● Convenient
● Extremely sensitive to low-copy target RNA
● Successful amplification up to 12 kb or more
● Reproducible RNA amplification

Description

HelixCript™ One-Step RT-PCR Kit is designed for sensitive amplification of the target gene in one-tube reaction from total transcripts. A reverse transcriptase, HelixCript™ Thermo Reverse Transcriptase, and a thermostable DNA polymerase, HelixAmp™ HyperSense-Taq polymerase, HelixAmp™ Speed-Pfu polymerase, or HelixAmp™ Hot-Taq polymeraseare supplied as an enzyme mixture. You can choose on of the enzymes according to your purpose.

One-Step RT-PCR Kit [HyperSense-Taq] : Long-range and high yield RT-PCR with low copy of target
One-Step RT-PCR Kit [Speed-Pfu] : Fast and high fidelity RT-PCR
One-Step RT-PCR Kit [Hot-Taq] : High-specific RT-PCR

One-step RT-PCR system provide the several advantages.
- Synthesis of cDNA and PCR amplification corresponding to target gene in one-tube reaction.
- Obtain the reproductive data in the repetitive experiment(s)
- Can save the time and cost for preparation of RT-PCR
- Amplification of low-copy transcripts by RT-PCR

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

Contents

Enzyme Mix, 2x Reaction Mix

 

Quality Control
The activity for cDNA synthesis and PCR amplification of target gene transcript using HelixCript™ One-Step RT-PCR Kit was evaluated by Limit-of Detection (LOD) assay and long range of gene constitutively-expressed in human total transcripts.

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

Figure 1. A fast RT-PCR by One-Step RT-PCR Kit [Speed-Pfu]. One-Step RT-PCR reactions were performed under the modified protocol as follow :The total time required in One-Step RT-PCR reaction is 1 hour. One-step RT-PCRs were performed with from 1 to 100 ng human blood total RNA using HelixCrip™ One-Step RT-PCR Kit [Speed-Pfu] and the primer set for human β-globin. PCR reaction was carried out 95℃, 10 sec and 55℃, 10 sec, 72℃, 20 sec (35 cycles).
Figure 2. Multiplex One-step RT-PCR of up to 4 targets by One-Step RT-PCR Kit [Hot-Taq]. Multiplex one-step RT-PCR in single-tube reaction was performed with total RNA extracted from hot pepper plant (Capsicum annuum) and by adding the indicated gene-specific primer sets. Each lane from 1 to 4 represents the progressive number of primer sets. The primer sets for the followed genes were used. CaAPX1 (374 bp), CaActin (461 bp), CaPF1 (615 bp), Cadhn (761 bp).
Figure 3. Viral RNA detection by One-Step RT-PCR Kit. Total RNA was isolated from a nasopharyngeal specimen of new influenza virus-infected patient and one-step RT-PCR was performed for the target corresponding to each influenza A (InfA, 106 bp), Swine influenza A (SW InfA, 195 bp), Swine Influenza H1 (SW H1,116 bp), RNase P (internal control, 65 bp)

Figure 4. Sensitivity test of One-Step RT-PCR Kit using Hepatitis A virus. One-step RT-PCR reactions were performed with purified HAV RNA and a primer set specific for HAV (191 bp) under the specified condition in protocol. Each PCR products were electrophoresed on 3% agarose gel. Lane 1: derived from 1/10 dilute of original sample; lane 2: 1/102 dilute; lane 3: 1/103 dilute; lane 4: 1/104 dilute; lane 5: 1/105 dilute; lane 6: 1/106 dilute; lane 7: 1/107 dilute; NTC: non-template control.
Cat.No. Product Size
 ORTHT100 HelixCript™ One-Step RT-PCR Kit [Hot-Taq] 100 rxns 
 ORTHT500 HelixCript™ One-Step RT-PCR Kit [Hot Taq] 500 rxns 
 ORTS100 HelixCript™ One-Step RT-PCR Kit [Speed-Pfu] 100 rxns 
 ORTS500 HelixCript™ One-Step RT-PCR Kit [Speed-Pfu] 500 rxns 
 ORTH100 HelixCript™ One-Step RT-PCR Kit [HyperSense-Taq] 100 rxns 
 ORTH500 HelixCript™ One-Step RT-PCR Kit [HyperSense-Taq] 500 rxns