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Visualization of Gene Expression by Fluorescent Multiplex Reverse Transcriptase-PCR Amplification

  • María Rosa Ponce
  • Víctor Quesada
  • Andrea Hricová
  • José Luis Micol
Part of the Methods in Molecular Biology book series (MIMB, volume 353)

Abstract

Many developmental and physiological analyses, population studies, and diagnostic tests can be performed by simply determining the presence or absence of a limited number of gene products. Here, we describe a rapid and sensitive procedure, based on the reverse transcription of total RNA samples followed by the co-amplification of specific complementary DNA molecules, for the simultaneous detection of different transcripts. Multiplex PCR amplification products are obtained in a single reaction mix containing several primer pairs, each of which includes a fluorescently labeled oligonucleotide; the amplification products are finally electrophoresed in an automated DNA sequencer controlled by fragment analysis software. The electropherograms obtained in this way allow a semiquantitative and efficient visualization of gene expression.

Key Words

Differential gene expression fluorescent labeling multiplex polymerase chain reaction transcription detection 

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Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • María Rosa Ponce
    • 1
    • 2
  • Víctor Quesada
    • 1
    • 2
  • Andrea Hricová
    • 1
    • 2
  • José Luis Micol
    • 1
    • 2
  1. 1.División de GenéticaUniversidad Miguel Hernández, Campus de ElcheAlicanteSpain
  2. 2.Instituto de BioingenieríaUniversidad Miguel Hernández, Campus de ElcheAlicanteSpain

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