[28] Reverse Transcription PCR (RT-PCR) Flashcards
What is Reverse Transcription PCR (RT-PCR)?
A laboratory technique combining reverse transcription of RNA into DNA and amplification of specific DNA targets via PCR.
What does RT-PCR stand for?
Reverse Transcription Polymerase Chain Reaction.
What is the key feature of RT-PCR?
It enables the conversion of RNA to DNA.
What are the steps involved in RT-PCR?
- RNA extraction
- Reverse transcription of RNA to cDNA
- PCR amplification of cDNA
- Detection of specific targets
What is the role of reverse transcription in RT-PCR?
Converts RNA into complementary DNA (cDNA).
What is RNA extraction?
The process of isolating RNA from a biological sample.
What is cDNA in the context of RT-PCR?
Complementary DNA synthesized from an RNA template through the process of reverse transcription.
What is the role of PCR in RT-PCR?
Amplifies specific DNA targets from the cDNA.
How is detection achieved in RT-PCR?
Through the identification of amplified DNA sequences, often using fluorescence.
What are the applications of RT-PCR?
- Virus detection
- Gene expression studies
- Molecular diagnostics
- Cancer research
Why is RT-PCR useful for virus detection?
Many viruses are RNA based. RT-PCR can convert this RNA into DNA for easier detection and study.
How is RT-PCR used in gene expression studies?
RT-PCR can quantify specific RNA levels, which can give insights into gene activity.
How does RT-PCR contribute to molecular diagnostics?
It provides a sensitive and accurate method for diagnosing diseases, particularly viral infections.
What is the role of RT-PCR in cancer research?
It helps in understanding the gene expression levels in different types of cancer, which can lead to potential treatments.
What makes RT-PCR a sensitive technique?
It can detect and quantify very small amounts of specific RNA sequences, even within complex mixtures.
What is the initial step in RT-PCR?
RNA extraction from the biological sample.
How is the cDNA synthesized in RT-PCR?
By using reverse transcriptase enzyme that uses RNA as a template to make cDNA.
What is PCR?
Polymerase Chain Reaction, a method widely used in molecular biology to make many copies of a specific DNA segment.
What is RNA?
Ribonucleic acid, a nucleic acid molecule similar to DNA but single-stranded and plays a vital role in coding, decoding, regulation, and expression of genes.
What is DNA?
Deoxyribonucleic acid, a molecule composed of two chains that coil around each other to form a double helix carrying genetic instructions.
