Reverse Transcriptase
Reverse Transcriptase - Related Videos
03:35
Assessing Viral Replication Capacity Using a Radiolabeled Reverse Transcriptase Assay
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2026
Source: Claiborne, D. T. et al. A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses. J. Vis. Exp. (2014)This video demonstrates the assessment of viral replication capacity using a radiolabeled reverse transcriptase assay. Culture supernatants containing progeny virions are added to a reverse transcription mixture containing a radiolabeled nucleotide. The incorporated signal reflects viral output and replication...
03:26
Reverse Transcriptase Activity as a Marker for Virus-Like Particle Production
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2026
Source: Scarborough, R. J., et al. Evaluation of the Efficacy And Toxicity of RNAs Targeting HIV-1 Production for Use in Gene or Drug Therapy. J. Vis. Exp. (2016).This video demonstrates a reverse transcriptase (RT) assay to assess the effect of a test RNA on virus-like particle (VLP) production. Cells are co-transfected with plasmids, and RT activity is measured as a quantitative indicator of VLP output. A decrease in RT activity reflects suppression of viral gene expression by the test RNA.
05:46
Rapid Screening of HIV Reverse Transcriptase and Integrase Inhibitors
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Cited by 15 •
2014
Here we describe cellular cytotoxicity and single round infectivity assays that allow for the rapid and accurate screening of compounds to determine their cellular cytotoxicity (CC50) and IC50 values against WT and drug resistant HIV-1.
04:52
Increasing cDNA Yields from Single-cell Quantities of mRNA in Standard Laboratory Reverse Transcriptase Reactions using Acoustic Microstreaming
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Cited by 11 •
2011
We describe a novel method for increasing cDNA yield from single-cell quantities of mRNA in otherwise standard laboratory reverse transcription reactions. The novelty resides in the use of a micromixer, which utilizes the phenomenon of acoustic microstreaming, to mix fluids at microliter scales more effectively than shaking, vortexing or trituration.
08:26
Semi-quantitative Detection of RNA-dependent RNA Polymerase Activity of Human Telomerase Reverse Transcriptase Protein
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Cited by 5 •
2018
Human telomerase reverse transcriptase (TERT) synthesizes not only telomeric DNA but also double-stranded RNA through RNA-dependent RNA polymerase activity. Here, we describe a newly established assay to detect RNA-dependent RNA polymerase activity of endogenous TERT.
FAQs
A primer gives reverse transcriptase a starting point for DNA synthesis on the RNA template. Once positioned, the enzyme uses available deoxynucleotides and base pairing to extend the complementary DNA strand. This requirement connects the RNA sequence to the resulting cDNA and makes primer selection an important part of experiments that examine particular RNA molecules.
RNase H activity degrades the RNA strand in an RNA-DNA product formed during reverse transcription. This activity is present in some reverse transcriptases and can affect how the RNA template is processed while cDNA is produced. Its presence therefore distinguishes enzyme activities that researchers may consider when selecting or interpreting a reverse-transcription system.
Base pairing allows the RNA template to guide the order of deoxynucleotides incorporated into the growing cDNA strand. The resulting DNA is complementary to the original RNA sequence rather than an independent copy with an unrelated sequence. This relationship enables researchers to use cDNA as a stable, analyzable representation of RNA information.
A typical reaction requires an RNA template, a primer, reverse transcriptase, and deoxynucleotides. The enzyme binds the RNA, the primer provides the initiation point, and the deoxynucleotides supply the building units for cDNA synthesis. Together, these components convert selected RNA information into DNA that can support downstream molecular biology analyses.
In reverse-transcription PCR, reverse transcriptase first converts RNA information into cDNA, creating a DNA form suitable for subsequent PCR analysis. This workflow links RNA molecules to an experimentally analyzable DNA product and supports studies of gene expression. It is especially useful when the biological material of interest is RNA rather than DNA.
cDNA construction preserves RNA-derived information in a DNA form that researchers can analyze and organize. It supports cDNA library construction and gene-expression analysis by converting RNA populations into complementary DNA collections or products. These applications allow investigators to examine which RNA information is present without analyzing the original RNA directly.
Reverse transcriptase provides a key biological connection between RNA genomes and DNA intermediates in retroviral replication. In research, the same activity also supports detection of RNA viruses by converting viral RNA into cDNA for analysis. Thus, the enzyme is relevant both to understanding retroviral biology and to identifying RNA-based infectious material.