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Analysis of PEDV Ag by Real-Time PCR Method (CAT#: STEM-MB-2909-LGZ)

Introduction

A real-time polymerase chain reaction (real-time PCR,or qPCR) is a laboratory technique of molecular biology based on the polymerase chain reaction(PCR). It monitors the amplification of a targeted DNA molecule during the PCR(i.e.,in real time),not at its end,as in conventional PCR.<br />Two common methods for the detection of PCR products in real-time PCR are(1)non-specific fluorescent dyes that intercalate with any double-stranded DNA and (2)sequence-specific DNA probes consisting of oligonucleotides that are labelled with a fluorescent reporter,which permits detection only after hybridization of the probe with its complementary sequence.




Principle

In this study, specific primers and probes were designed for highly conserved regions of porcine epidemic diarrhea virus. PCR reaction was performed and fluorescence signals were released when the response system contained porcine epidemic diarrhea virus genomic template. The fluorescence quantitative PCR instrument was used for real-time monitoring and output of the corresponding channel signals in the process of PCR to achieve qualitative analysis of the detection results.

Applications

Qualitative detection of porcine epidemic diarrhea virus, caused to the pig epidemic diarrhea virus disease diagnosis and efficacy evaluation has important guiding significance.

Procedure

1. Sample processing and preparation of PCR reaction system.
2. Add the amplification template, cover the PCR reaction cover, mix well, centrifuge at low speed instantaneously, and transfer to the PCR instrument.
3. Set the program for PCR amplification.
4. Data analysis.

Materials

• Sample Type: Serum/intestinal contents/tissue organs