Comparison of whole-genome amplifications for microsatellite genotyping of Rotylenchulus reniformis

Currently, a large number of microsatellites are available for Rotylenchulus reniformis (reniform nematode); however, two barriers exist for genotyping samples from different geographical areas. The limited amount of nucleic acids obtained from single nematodes which would require their multiplicati...

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Autores principales:	Arias,Renée S, Stetina,Salliana R, Scheffler,Brian E
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2011
Materias:	genotyping reniform nematode SSR STR WGA
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000300014
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spelling	oai:scielo:S0717-345820110003000142011-09-23Comparison of whole-genome amplifications for microsatellite genotyping of Rotylenchulus reniformisArias,Renée SStetina,Salliana RScheffler,Brian E genotyping reniform nematode SSR STR WGA Currently, a large number of microsatellites are available for Rotylenchulus reniformis (reniform nematode); however, two barriers exist for genotyping samples from different geographical areas. The limited amount of nucleic acids obtained from single nematodes which would require their multiplication to obtain enough DNA for testing; and the strictly regulated transport of live samples and multiplication in greenhouse for being a plant pathogen. Whole-genome amplification (WGA) of samples consisting of one and five dead gravid females with their associated egg masses was successfully performed on disrupted tissue using three commercial kits. DNA yield after WGA ranged from 0.5 to 8 Âµg and was used to test 96 microsatellite markers we previously developed for the reniform nematode. The results were compared to those of fingerprinting the original population (MSRR03). Out of 96 markers tested, 71 had amplicons in MSRR03. Using WGA of single gravid females with their associated egg masses, 86-93% of the alleles found on MSRR03 were detected, and 87-88% of the alleles found on MSRR03 when using WGA of samples composed of five gravid females with their associated egg masses as template. Our results indicate that reniform nematode samples as small as a single gravid female with her associated egg mass can be used in WGA and direct testing with microsatellites, giving consistent results when compared to the original population.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.14 n.3 20112011-05-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000300014en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	genotyping reniform nematode SSR STR WGA
spellingShingle	genotyping reniform nematode SSR STR WGA Arias,Renée S Stetina,Salliana R Scheffler,Brian E Comparison of whole-genome amplifications for microsatellite genotyping of Rotylenchulus reniformis
description	Currently, a large number of microsatellites are available for Rotylenchulus reniformis (reniform nematode); however, two barriers exist for genotyping samples from different geographical areas. The limited amount of nucleic acids obtained from single nematodes which would require their multiplication to obtain enough DNA for testing; and the strictly regulated transport of live samples and multiplication in greenhouse for being a plant pathogen. Whole-genome amplification (WGA) of samples consisting of one and five dead gravid females with their associated egg masses was successfully performed on disrupted tissue using three commercial kits. DNA yield after WGA ranged from 0.5 to 8 Âµg and was used to test 96 microsatellite markers we previously developed for the reniform nematode. The results were compared to those of fingerprinting the original population (MSRR03). Out of 96 markers tested, 71 had amplicons in MSRR03. Using WGA of single gravid females with their associated egg masses, 86-93% of the alleles found on MSRR03 were detected, and 87-88% of the alleles found on MSRR03 when using WGA of samples composed of five gravid females with their associated egg masses as template. Our results indicate that reniform nematode samples as small as a single gravid female with her associated egg mass can be used in WGA and direct testing with microsatellites, giving consistent results when compared to the original population.
author	Arias,Renée S Stetina,Salliana R Scheffler,Brian E
author_facet	Arias,Renée S Stetina,Salliana R Scheffler,Brian E
author_sort	Arias,Renée S
title	Comparison of whole-genome amplifications for microsatellite genotyping of Rotylenchulus reniformis
title_short	Comparison of whole-genome amplifications for microsatellite genotyping of Rotylenchulus reniformis
title_full	Comparison of whole-genome amplifications for microsatellite genotyping of Rotylenchulus reniformis
title_fullStr	Comparison of whole-genome amplifications for microsatellite genotyping of Rotylenchulus reniformis
title_full_unstemmed	Comparison of whole-genome amplifications for microsatellite genotyping of Rotylenchulus reniformis
title_sort	comparison of whole-genome amplifications for microsatellite genotyping of rotylenchulus reniformis
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2011
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000300014
work_keys_str_mv	AT ariasrenees comparisonofwholegenomeamplificationsformicrosatellitegenotypingofrotylenchulusreniformis AT stetinasallianar comparisonofwholegenomeamplificationsformicrosatellitegenotypingofrotylenchulusreniformis AT schefflerbriane comparisonofwholegenomeamplificationsformicrosatellitegenotypingofrotylenchulusreniformis
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Comparison of whole-genome amplifications for microsatellite genotyping of Rotylenchulus reniformis

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