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Related Concept Videos

MicroRNAs01:22

MicroRNAs

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
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High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
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A high-throughput microRNA expression profiling system.

Yanwen Guo1, Stephen Mastriano, Jun Lu

  • 1Department of Genetics, Yale Stem Cell Center, Yale University, 10 Amistad Street, New Haven, CT, 06520-8005, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 18, 2014
PubMed
Summary
This summary is machine-generated.

This study presents a new, cost-effective method for high-throughput microRNA (miRNA) expression profiling. The robust protocol enables simultaneous analysis of hundreds of samples, aiding diagnostic and prognostic research.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • MicroRNAs (miRNAs) are small noncoding RNAs regulating biological functions.
  • miRNA expression levels offer diagnostic, prognostic, and pharmacologic response insights.
  • Large-scale miRNA expression profiling is crucial for advancing research.

Purpose of the Study:

  • To introduce a robust, high-throughput protocol for miRNA expression profiling.
  • To enable simultaneous analysis of hundreds of samples for miRNA detection.
  • To provide a flexible and cost-effective solution for miRNA research.

Main Methods:

  • Utilizes 96-well-based microRNA capturing from total RNA.
  • Incorporates on-site biochemical reactions for sample processing.
  • Employs bead-based detection in a 96-well format for comprehensive miRNA analysis.

Main Results:

  • The protocol supports high-throughput sample labeling and detection.
  • Enables profiling of hundreds of miRNAs per sample simultaneously.
  • Demonstrates low-cost, high specificity, and flexibility for various sample sizes.

Conclusions:

  • The developed protocol is suitable for widespread laboratory adoption.
  • It facilitates efficient and accurate global miRNA expression profiling.
  • This method supports large-scale studies in diagnostics and prognostics.