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Analysis of Canine Adenovirus Type I (CAV-I) by Real-Time PCR Method (CAT#: STEM-MB-2553-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.
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

Specific primers and probes were designed for highly conserved regions of the canine adenovirus type I genome. PCR reaction was performed and fluorescence signals were released when the canine adenovirus type I genome template was included in the reaction system. Real-Time PCR instrument was used to monitor and output the corresponding channel signals in the process of PCR, and the qualitative analysis of detection results was realized.

Applications

It is suitable for canine adenovirus type I gene test and can be used for auxiliary diagnosis of clinical canine adenovirus type I 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: serum, plasma, tissue
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