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Analysis of Mycoplasma Spp. by Real-Time PCR Method (CAT#: STEM-MB-2877-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

Mycoplasmosomes are the smallest and simplest prokaryotes ever discovered. Their lack of cell walls, ability to deform, ability to pass through filters, and ability to grow and reproduce in non-living culture-medium have become a common source of pollution in cell culture, seriously affecting cell growth rate, cell morphology, gene expression, cell metabolism and cell viability. Therefore, rapid and sensitive detection of mycoplasma is of great significance. Primers and probes involved in this experiment have been optimized with high sensitivity. Primers are highly specific. Primers are designed according to the highly conserved region of Mycoplasma and will not cross-react with the DNA of other microorganisms.

Applications

It is used for rapid and sensitive detection of mycoplasma.

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
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