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Related Concept Videos

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...
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...
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...

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Electrophoretic Mobility Shift Assay (EMSA) for the Study of RNA-Protein Interactions: The IRE/IRP Example
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Electrical sensor array for polymerase chain reaction-free messenger RNA expression profiling.

Xiaojun Chen1, Somenath Roy, Yanfen Peng

  • 1Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, Singapore 138669.

Analytical Chemistry
|June 29, 2010
PubMed
Summary
This summary is machine-generated.

This study introduces a novel electrical sensor array for sensitive messenger RNA (mRNA) expression profiling without polymerase chain reaction (PCR) amplification. The device achieves ultrasensitive mRNA detection and discrimination using a unique nanogap structure and hybridization technique.

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Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays

Published on: December 3, 2010

Area of Science:

  • Biotechnology
  • Electrical Engineering
  • Molecular Biology

Background:

  • Messenger RNA (mRNA) expression profiling is crucial for understanding cellular function and disease.
  • Current methods often rely on polymerase chain reaction (PCR) amplification, which can introduce biases and increase complexity.
  • There is a need for simple, sensitive, and accurate methods for direct mRNA detection and quantification.

Purpose of the Study:

  • To develop a novel electrical sensor array for direct and sensitive mRNA expression profiling.
  • To demonstrate the capability of the sensor array for ultrasensitive mRNA detection without PCR amplification.
  • To evaluate the sensor array's performance in terms of sensitivity, linearity, and mismatch discrimination.

Main Methods:

  • Fabrication of a silicon chip with vertically aligned gold microband electrode/SiO(2)/gold microband electrode sandwich structures (nanogap sensors).
  • Two-step hybridization strategy: target mRNA hybridized with capture probes (CP) on the bottom, and its poly(A) tail with annealing probes (AP) on the top, vertically aligning mRNA across the nanogap.
  • Subsequent metallization of hybridized mRNA to bridge the nanogap, enabling conductance-based detection.

Main Results:

  • Ultrasensitive detection of mRNA down to 0.10 femtomolar (fM) was achieved.
  • A linear relationship between conductance and mRNA concentration was observed from 0.50 fM to 1.0 picomolar (pM).
  • The sensor array successfully detected as little as a 50% difference in mRNA expression and showed excellent mismatch discrimination.

Conclusions:

  • The developed electrical sensor array offers a simple, sensitive, and PCR-free platform for mRNA expression profiling.
  • The unique nanogap structure and two-step hybridization enable ultrasensitive detection and high specificity.
  • This technology has the potential for various applications in molecular diagnostics and biological research.