In Reverse Transcription PCR, what step is added the ordinary PCR Reaction?
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Answer:
REVERSE TRANSCRIPTION–POLYMERASE CHAIN REACTION
RT-PCR uses RNA as starting material for in vitro nucleic acid amplification. The discovery of retroviral reverse transcriptase in the early 1970s ultimately made RT-PCR possible. Reverse transcriptase is an RNA-dependent DNA polymerase, catalyzing DNA synthesis using RNA as the template. The end product is known as complementary DNA (cDNA). cDNA is not subject to RNase degradation, making it more stable than RNA. In RT-PCR, the starting RNA is subsequently degraded, dsDNA is produced, and PCR amplification proceeds in the usual manner. RNA extraction kits for both manual and automated RNA purification exist and, when combined with RT-PCR, make RNA analysis in the clinical laboratory virtually as rapid and equally sensitive as PCR-based DNA amplification. RT-PCR is commonly used in the diagnosis and quantification of RNA virus infections (e.g., human immunodeficiency virus and hepatitis C virus) and the analysis of mRNA transcripts such as those produced by translocations commonly associated with non-Hodgkin's lymphomas, leukemias, and sarcomas. Gene expression profiling is likely to have a major impact on molecular diagnostics in the coming years and will depend on RNA analysis using RT-PCR and possibly high-density arrays.
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