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Related Concept Videos

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Reporter Genes

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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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Transcription Start Site Mapping Using Super-low Input Carrier-CAGE
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Detecting expressed genes using CAGE.

Mitsuyoshi Murata1, Hiromi Nishiyori-Sueki, Miki Kojima-Ishiyama

  • 1Division of Genomic Technologies, RIKEN Center for Life Science Technologies, RIKEN Yokohama Institute, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|June 15, 2014
PubMed
Summary
This summary is machine-generated.

We developed a new Cap analysis of gene expression (CAGE) method called nAnT-iCAGE. This protocol reduces bias in RNA expression analysis by excluding PCR amplification and enzyme cleavage, simplifying library preparation.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Transcriptomics

Background:

  • Cap analysis of gene expression (CAGE) is a high-throughput method for measuring RNA expression.
  • CAGE analysis of transcript 5' ends aids in determining transcription start sites and predicting promoter regions.

Purpose of the Study:

  • To provide a protocol for constructing no-amplification non-tagging CAGE libraries (nAnT-iCAGE).
  • To reduce bias in CAGE library preparation for Illumina sequencers.

Main Methods:

  • Developed the nAnT-iCAGE protocol, excluding PCR amplification and restriction enzyme cleavage.
  • Applied the protocol for constructing CAGE libraries compatible with Illumina next-generation sequencing.

Main Results:

  • Achieved a less biased and simpler CAGE library preparation process.
  • Enabled accurate high-throughput measurement of RNA expression with reduced potential biases.

Conclusions:

  • The nAnT-iCAGE protocol offers a streamlined and less biased approach for CAGE library construction.
  • This method enhances the reliability of RNA expression profiling and promoter analysis.