Reverse Transcription

Reverse transcription is the biological process of synthesizing DNA from an RNA template, reversing the usual flow of genetic information and enabling researchers to study RNA molecules through stable DNA copies. The enzyme reverse transcriptase binds RNA, uses a primer to initiate synthesis, and incorporates complementary DNA nucleotides to produce a complementary DNA (cDNA) strand. In biology, reverse transcription is central to the replication of retroviruses and supports laboratory methods such as reverse transcription polymerase chain reaction (RT-PCR), cDNA library construction, and gene-expression analysis. These applications help quantify transcripts and investigate cellular responses, development, and disease.

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JoVE EoE - PCR Techniques

Real-Time Quantitative Reverse Transcription PCR for Diagnosing Viral Infections

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2025

This video demonstrates a quantitative method for detecting viral infection using real-time reverse transcription PCR. The reverse transcription step converts the RNA to cDNA, which is then amplified via PCR cycles. The presence of viral RNA in the sample is confirmed by analyzing the amplification and dissociation curves of DNA obtained from the PCR.

Measuring Hepatitis C Viral Load Using Quantitative Reverse Transcription Polymerase Chain Reaction

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2026

Source: Ren, S., et al. A Protocol for Analyzing Hepatitis C Virus Replication. J. Vis. Exp. (2014).This video demonstrates the procedure for quantifying Hepatitis C viral RNA using quantitative reverse transcription PCR to measure viral load in infected cell cultures.

High Throughput In Vitro Assessment of Latency Reversing Agents on HIV Transcription and Splicing

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2019

A high throughput protocol for functional assessment of HIV efficient reactivation and clearance of latent proviruses is described and applied by testing the impact of interventions on HIV transcription and splicing. Representative results of the effect of latency reversing agents on LTR-driven transcription and splicing are provided.

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp

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Cited by 15 •

2018

This protocol describes the critical steps and precautions required to perform single cell multiplex reverse transcription polymerase chain reaction after patch-clamp. This technique is a simple and effective method to analyze the expression profile of a predetermined set of genes from a single cell characterized by patch-clamp recordings.

Analysis of Termination of Transcription Using BrUTP-strand-specific Transcription Run-on (TRO) Approach

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Cited by 10 •

2017

We describe a basic experimental approach for analysis of termination of transcription by RNA polymerase II in vivo using BrUTP by the strand-specific transcription run-on (TRO) approach in budding yeast. This protocol can be extended to study transcription termination by other RNA polymerases both in yeast and higher eukaryotes.

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