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Related Concept Videos

RNA-seq03:21

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
Next-generation Sequencing03:00

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
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Ribosome Profiling02:24

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
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RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer.
Since the...
Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
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Related Experiment Video

Updated: Jun 17, 2026

Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations
11:52

Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations

Published on: August 4, 2016

FRT-seq: amplification-free, strand-specific transcriptome sequencing.

Lira Mamanova1, Robert M Andrews, Keith D James

  • 1The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.

Nature Methods
|January 19, 2010
PubMed
Summary
This summary is machine-generated.

This study introduces a novel transcriptome sequencing method for Illumina Genome Analyzer. The new approach avoids PCR biases and duplicates, yielding strand-specific sequences directly from poly(A)(+) RNA.

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Last Updated: Jun 17, 2026

Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations
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07:27

Transcriptome Analysis of Single Cells

Published on: April 25, 2011

Area of Science:

  • Molecular Biology
  • Genomics
  • Next-Generation Sequencing

Background:

  • Traditional transcriptome sequencing methods often involve PCR amplification, which can introduce biases and duplicate reads.
  • Strand specificity is crucial for accurate gene expression analysis, but not always achieved with standard protocols.

Purpose of the Study:

  • To develop an alternative library preparation method for transcriptome sequencing on the Illumina Genome Analyzer.
  • To eliminate PCR amplification biases and generate strand-specific sequencing data.
  • To ensure compatibility with both paired-end and single-end sequencing.

Main Methods:

  • Reverse transcription is performed directly on the flowcell.
  • Poly(A)(+) RNA is used as the template, bypassing cDNA synthesis and amplification steps.
  • The method is compatible with standard Illumina sequencing platforms.

Main Results:

  • Elimination of PCR biases and duplicate reads.
  • Generation of inherently strand-specific sequencing data.
  • Successful application with both paired-end and single-end sequencing strategies.

Conclusions:

  • This flowcell-based reverse transcription method offers a streamlined and unbiased approach to transcriptome sequencing.
  • The resulting strand-specific data enhances the accuracy of gene expression studies.
  • The method provides a valuable alternative for researchers using Illumina sequencing technology.