Use of an intron containing grapevine gene as internal control for validation of cDNA synthesis in virus detection by RT-PCR

Róbert Oláh, Tamás Deák, Mihály Turcsán, Márta Szénási, Ádám Bordé, E. Szegedi

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Detection of viruses in grapevine plants is important to prevent their spreading with propagating stocks. For this purpose PCR based protocols are commonly and routinely used. Since most grapevine viruses are RNA viruses, the detection procedure starts with RNA purification followed by cDNA synthesis prior to PCR. The cDNAs should be validated by an internal control for which usually constitutively expressed housekeeping genes are used. Here we publish a new set of primers designed in the Vitis vinifera phosphoenolpyruvate carboxylase gene to encompass two or three introns. PCR products amplified from remaining genomic DNA in the RNA samples and from cDNA can be clearly distinguished since cDNA-derived products are smaller in size compared to those amplified from genomic DNA. Thus these primers may be useful reference markers in RT-PCR-based virus detection assays both for the validation of cDNA synthesis and the detection of trace amounts of genomic DNA.

Original languageEnglish
Pages (from-to)765-770
Number of pages6
JournalEuropean Journal of Plant Pathology
Volume149
Issue number3
DOIs
Publication statusPublished - Nov 1 2017

Fingerprint

introns
reverse transcriptase polymerase chain reaction
viruses
synthesis
genomics
DNA
genes
RNA
phosphoenolpyruvate carboxylase
Vitis vinifera
assays
sampling

Keywords

  • cDNA
  • Polymerase chain reaction
  • Virus detection
  • Vitis spp

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science
  • Horticulture

Cite this

Use of an intron containing grapevine gene as internal control for validation of cDNA synthesis in virus detection by RT-PCR. / Oláh, Róbert; Deák, Tamás; Turcsán, Mihály; Szénási, Márta; Bordé, Ádám; Szegedi, E.

In: European Journal of Plant Pathology, Vol. 149, No. 3, 01.11.2017, p. 765-770.

Research output: Contribution to journalArticle

Oláh, Róbert ; Deák, Tamás ; Turcsán, Mihály ; Szénási, Márta ; Bordé, Ádám ; Szegedi, E. / Use of an intron containing grapevine gene as internal control for validation of cDNA synthesis in virus detection by RT-PCR. In: European Journal of Plant Pathology. 2017 ; Vol. 149, No. 3. pp. 765-770.
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