Advertisement

Planta

, Volume 211, Issue 4, pp 606–608 | Cite as

A multiplex reverse transcriptase-polymerase chain reaction method for fluorescence-based semiautomated detection of gene expression in Arabidopsis thaliana

  • María Rosa Ponce
  • José Manuel Pérez-Pérez
  • Pedro Piqueras
  • José Luis Micol
Rapid communication

Abstract.

 A non-radioactive, rapid and sensitive method is presented for the simultaneous detection of several mRNA molecules. The technique is based on conventional first-strand cDNA synthesis by reverse transcriptase, followed by multiplex polymerase chain reaction (PCR) co-amplification of several gene products in a reaction mix containing several primer sets, each including a fluorescently labeled oligonucleotide. The PCR products obtained are finally electrophoresed in a single lane of a polyacrylamide gel, in an automated DNA sequencer controlled by fragment-analysis software. The method has proven useful to efficiently detect nine mRNA transcripts, some of which are low copy number, from small specimens such as single flowers and leaves of Arabidopsis thaliana (L.) Heynh. This approach might be easily extended to other biological systems, for developmental and physiological analyses, population studies and diagnosis.

Key words: Arabidopsis– Differential gene expression – Multiplex polymerase chain reaction – Transcription detection 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Copyright information

© Springer-Verlag Berlin Heidelberg 2000

Authors and Affiliations

  • María Rosa Ponce
    • 1
  • José Manuel Pérez-Pérez
    • 1
  • Pedro Piqueras
    • 1
  • José Luis Micol
    • 1
  1. 1.División de Genética, Universidad Miguel Hernández, Campus de San Juan, 03550 Alicante, SpainES

Personalised recommendations