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TaqMan Array Cards in pharmaceutical research.

David N Keys1, Janice K Au-Young, Richard A Fekete

  • 1Molecular Biology Division, Life Technologies Corporation, Foster City, CA, USA. david.keys@lifetech.com

Methods in Molecular Biology (Clifton, N.J.)
|March 11, 2010
PubMed
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TaqMan Array Cards offer accurate, high-throughput gene expression analysis for mRNA and miRNA. This protocol streamlines transcription assays in cultured cells by skipping RNA isolation for faster results.

Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Quantitative analysis of gene expression is crucial in biological research.
  • Real-time PCR (Polymerase Chain Reaction) is a standard method for transcript quantification.
  • High-throughput methods are needed to analyze multiple genes simultaneously.

Purpose of the Study:

  • To describe a protocol for high-throughput transcription analysis using TaqMan Array Cards.
  • To optimize the protocol for cultured cells, minimizing hands-on time and pipetting.
  • To enable direct cDNA generation from cell lysates, bypassing RNA isolation.

Main Methods:

  • Utilizing microfluidic TaqMan Array Cards with 384 reaction chambers.
  • Employing Applied Biosystems TaqMan Gene Expression Assays for real-time PCR monitoring.

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  • Implementing a protocol that skips RNA isolation and generates cDNA directly in lysis solution.

    • Main Results:

    • TaqMan Array Cards provide accurate, sensitive, and simple quantitative analysis of mRNA or miRNA.
    • The described protocol minimizes hands-on time and pipetting steps.
    • Direct cDNA generation in lysis solution is feasible for transcription assays.

    Conclusions:

    • TaqMan Array Cards are effective tools for high-throughput gene expression studies.
    • The optimized protocol offers an efficient method for analyzing transcription in cultured cells.
    • This approach facilitates verification of microarray data and hypothesis-driven gene panel testing.