Efficiency of Real-Time PCR for 18s rRNA amplification of Sorbus domestica L.

Authors

  • Jana Žiarovská Department of Genetics and Plant Breeding SAU in Nitra Tr. A. Hlinku 94976 Nitra
  • Petronela Poláčeková Slovak University of Agriculture, Faculty of Agrobiology and Food Resources, Department of Plant Genetics and Breeding, Tr. A. Hlinku 2, 949 76 Nitra

DOI:

https://doi.org/10.5219/203

Keywords:

Sorbus domestica, L., 18S rRNA, real-time PCR

Abstract

Nowadays, the awareness is given more and more to underutilized and  unusual fruits. One of them is Sorbus domestica, L. not only as an endangered species, but as well as a promising and economically usable crop. The work was aimed for finding a total genomic DNA isolating methods from fresh plant material and confirmation of the optimized method by the detection of 18S rRNA gene using real-time PCR. Two commercial isolation kits were tested -  Invisorb® Spin Plant Mini Kit and Wizard ® Genomic DNA. Higher purity and yield of DNA isolation kit showed Invisorb kit. The effective and pure PCR amplification was confirmed for Invisorb, too when 20 ng undiluted DNA at annealing temperature of 64.5 °C.

Downloads

Download data is not yet available.

Metrics

Metrics Loading ...

References

BAJZÍK, P., GOLIAN, J., ŽIDEK, R., ČAPLA, J., BELEJ, Ľ., ONDREJKA, M., MRÁZOVÁ, Ľ., MARŠÁLKOVÁ, L. 2010. Methods for fish species identification in foodproducts, In Potravinarstvo, vol. 4, 2010, no. 3, p. 1-5. https://doi.org/10.5219/25 DOI: https://doi.org/10.5219/25

BAJZÍK, P., ŽIDEK, R., GOLIAN, J., BELEJ, Ľ., ČAPLA, J., MARŠÁLKOVÁ, L., REVÁK, O. Optimalisation of species identification of common carp (Cyprinus carpio) using SYBR® GREEN I real-time PCR method. In Potravinarstvo, vol. 5, 2011, no. 3, p. 1-5. https://doi.org/10.5219/154 DOI: https://doi.org/10.5219/154

BENALI, S., MOHAMED, B., EDDINE, H.J., NEEMA, C. 2011. Advances of Molecular Markers Application in Plant Pathology Research. In European Journal of Scientific Research. vol.50, 2011, no.1, pp.110-123.

BRINDZA, J., ČERVEŇÁKOVÁ, J., TÓTH, D., BÍRO, D., ŠAJBIDOR, J. 2009. Unutilized potential of True Service Tree (Sorbus domestica, L.) In Acta Hort. (ISHS) vol. 806, 2009, p. 717-726. DOI: https://doi.org/10.17660/ActaHortic.2009.806.89

LIN, J.J., FLEMING, R., KUO, J., MATTHEWS, B.F., SAUNDERS, J.A. 2000. Detection of Plant Genes Using a Rapid, Nonorganic DNA Purification Method. In BioTechniques,vol. 28, 2000, p. 346-350. PMiD: 10683746 DOI: https://doi.org/10.2144/00282pf01

MEYER, A., TODT, CH., MIKKELSEN, N. T., LIEB, B. 2010. Fast evolving 18S rRNA sequences from Solenogastres (Mollusca) resist standard PCR amplification and give new insights into mollus substitution rate heterogeneity. In BMC EvolutionaryBiology. 2010, vol. 10. https://doi.org/10.1186/1471-2148-10-70, PMiD: 20214780 DOI: https://doi.org/10.1186/1471-2148-10-70

REISCHL, U., WITTWER, C.T., COCKERILL, F. 2002. Rapid Cycle Real-time PCR: Methods and Applications; Microbiology and FoodAnalysis. New York Springer-Verlag, 2002. https://doi.org/10.1007/978-3-642-48351-6_8,

ŠKULTÉTY, O., JURČÁKOVÁ, A. 2011. EVA GREEN real-time PCR used to detect cellery as an allergen in food, In Potravinarstvo, vol. 5, 2011, no. 2, p. 70-72.https://doi.org/ 10.5219/138 DOI: https://doi.org/10.5219/138

WITTWER, C.T., RIRIE, K.M., ANDREW, R.V. et al. 1997. The Light Cycler: a microvolume multisample fluorimeter with rapid temperature control. In Biotechniques, vol. 22, 1997, p. 176-181. PMiD: 8994665 DOI: https://doi.org/10.2144/97221pf02

WONG, M.L., MEDRANO, J.F. 2005. Real-time PCR form RNA quantitation. In BioTechniquesvol. 39, 2005, p. 75-85. https://doi.org/10.2144/05391RV01 DOI: https://doi.org/10.2144/05391RV01

ZELEŇÁKOVÁ, L., ŽIDEK, R., ČANIGOVÁ, M.. PAULOV, J., GALLISOVÁ, T. 2010. Evaluation of ELISA method to detection of cowβ-lactoglobulin in sheepmilk and sheepmilk products, In Potravinarstvo, vol. 4, 2010, no. 4, p. 80-84. https://doi.org/10.5219/78 DOI: https://doi.org/10.5219/78

ŽIAROVSKÁ, J., RAŽNÁ, K., SENKOVÁ, S., ŠTEFÚNOVÁ, V., BEŽO M. Variability of Linum usitatissimum L. based on molecular markers. In ARPN Journal of Agricultural and Biological Science,vol. 7, 2012, p. 50-58.

Downloads

Published

2012-10-11

How to Cite

Žiarovská, J. ., & Poláčeková, P. . (2012). Efficiency of Real-Time PCR for 18s rRNA amplification of Sorbus domestica L. Potravinarstvo Slovak Journal of Food Sciences, 6(3), 47–49. https://doi.org/10.5219/203