QRT-PCR

Quantitative Reverse Transcription Polymerase Chain Reaction (QRT-PCR), also known as Real-Time Reverse Transcription PCR or RT-qPCR, is a laboratory technique used in molecular biology to amplify and simultaneously quantify a targeted DNA molecule. It enables both detection and quantification (as absolute number of copies or relative amount when normalized to DNA input or additional normalizing genes) of a specific sequence in a DNA sample.

Overview

The QRT-PCR technique is based on the polymerase chain reaction (PCR), which is used to amplify a specific DNA sequence. Unlike the traditional PCR, QRT-PCR is able to amplify and simultaneously quantify the targeted DNA molecule. This is achieved by the fluorescent reporter, which emits fluorescence in real-time as the reaction progresses, allowing the quantification of the DNA.

Procedure

The procedure of QRT-PCR involves the reverse transcription of the RNA into complementary DNA (cDNA) using the enzyme reverse transcriptase. This is followed by amplification of the cDNA using PCR. The quantification of the DNA is achieved by the fluorescent reporter, which emits fluorescence in real-time as the reaction progresses.

Applications

QRT-PCR is widely used in research for gene expression analysis, genetic variation analysis, genotyping, and genetically modified organism (GMO) detection. It is also used in clinical diagnostics and forensic science.

Advantages and Limitations

The main advantage of QRT-PCR is its ability to amplify and simultaneously quantify a specific DNA sequence. However, it requires a high level of expertise and expensive equipment. It is also sensitive to contamination and requires careful sample preparation.

See Also

• Polymerase chain reaction
• Molecular biology
• DNA
• RNA
• cDNA
• Gene expression
• Genetic variation
• Genotyping
• Genetically modified organism
• Clinical diagnostics
• Forensic science

QRT-PCR Resources
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