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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...
DNA Microarrays02:34

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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.
The real-time quantification of the number of amplified products is...
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...

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MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
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MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method

Published on: October 7, 2025

Microarray-based amplification and detection of RNA by nucleic acid sequence based amplification.

Andreas Mader1, Ulrike Riehle, Thomas Brandstetter

  • 1Institute of Pathology, Freiburg University Medical Center, Breisacher Strasse 115a, 79106 Freiburg, Germany.

Analytical and Bioanalytical Chemistry
|July 3, 2010
PubMed
Summary

This study introduces a one-step RNA amplification and labeling method using Nucleic Acid Sequence-Based Amplification (NASBA) combined with microarray analysis. This novel approach enables sensitive detection of breast cancer biomarkers, potentially allowing for single-cell assays.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genomics

Background:

  • Nucleic Acid Sequence-Based Amplification (NASBA) is a powerful in vitro nucleic acid amplification technique.
  • Microarray analysis is widely used for gene expression profiling.

Purpose of the Study:

  • To develop a one-step process combining NASBA and microarray analysis for RNA amplification and labeling.
  • To demonstrate the feasibility of multiplex NASBA-on-microarray analysis for breast cancer diagnostics.

Main Methods:

  • A one-step process integrating NASBA with microarray analysis was developed.
  • NASBA reactions were performed in direct contact with capture probes immobilized on hydrogel spots on chip surfaces.
  • A printing and UV irradiation process was used to generate hydrogel spots on chip surfaces.

Main Results:

  • Five gene expression and SNP parameters relevant to breast cancer diagnostics were successfully analyzed.
  • A minimum of 10 pg of total RNA was sufficient for detecting the RPS18 reference parameter.
  • The developed NASBA-on-microarray assay demonstrated high sensitivity, with potential for single-cell analysis.

Conclusions:

  • Multiplex NASBA-on-microarray analysis is a feasible and sensitive method for RNA detection.
  • The assay shows promise for breast cancer diagnostics and molecular profiling.
  • The high sensitivity achieved suggests potential applications in single-cell analysis.