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Analysis of NDV Ag by Real-Time PCR Method (CAT#: STEM-MB-2896-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 experiment, specific primers and probes were designed for highly conserved regions of NDV Ag genome. PCR reaction was carried out and fluorescence signals were released under the condition that NDV Ag genomic template was included in the reaction system. 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

It is suitable for NDV Ag detection, and can be used for auxiliary diagnosis of clinical NDV Ag infection, but not for diagnosis.

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: Throat swab/cloaca swab/blood/poultry muscle/viscera