Application of PCR-HRM method for microsatellite polymorphism genotyping in the LDHA gene of pigeons (Columba livia).

High-resolution melting (HRM) is a post-PCR method that allows to discriminate genotypes based on fluorescence changes during the melting phase. HRM is used to detect mutations or polymorphisms (e.g. microsatellites, SNPs, indels). Here, the (TTTAT)3-5 microsatellite polymorphism within intron 6 of...

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Autores principales:	Magdalena Jedrzejczak-Silicka, Adam Lepczynski, Filip Gołębiowski, Daniel Dolata, Andrzej Dybus
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4587776b885e4fddb81d15611b2eb08f
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spelling	oai:doaj.org-article:4587776b885e4fddb81d15611b2eb08f2021-12-02T20:17:46ZApplication of PCR-HRM method for microsatellite polymorphism genotyping in the LDHA gene of pigeons (Columba livia).1932-620310.1371/journal.pone.0256065https://doaj.org/article/4587776b885e4fddb81d15611b2eb08f2021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0256065https://doaj.org/toc/1932-6203High-resolution melting (HRM) is a post-PCR method that allows to discriminate genotypes based on fluorescence changes during the melting phase. HRM is used to detect mutations or polymorphisms (e.g. microsatellites, SNPs, indels). Here, the (TTTAT)3-5 microsatellite polymorphism within intron 6 of the LDHA gene in pigeons was analysed using the HRM method. Individuals (123 homing pigeons) were genotyped using conventional PCR. Birds were classified into groups based on genotype type and the results were tested by qPCR-HRM and verified using sequencing. Based on the evaluated protocol, five genotypes were identified that vary in the number of TTTAT repeat units (3/3, 4/4, 3/4, 4/5, and 5/5). Sequencing have confirmed the results obtained with qPCR-HRM and verified that HRM is a suitable method for identification of three-allele microsatellite polymorphisms. It can be concluded that the high-resolution melting (HRM) method can be effectively used for rapid (one-step) discrimination of the (TTTAT)3-5 microsatellite polymorphism in the pigeon's LDHA gene.Magdalena Jedrzejczak-SilickaAdam LepczynskiFilip GołębiowskiDaniel DolataAndrzej DybusPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 8, p e0256065 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Magdalena Jedrzejczak-Silicka Adam Lepczynski Filip Gołębiowski Daniel Dolata Andrzej Dybus Application of PCR-HRM method for microsatellite polymorphism genotyping in the LDHA gene of pigeons (Columba livia).
description	High-resolution melting (HRM) is a post-PCR method that allows to discriminate genotypes based on fluorescence changes during the melting phase. HRM is used to detect mutations or polymorphisms (e.g. microsatellites, SNPs, indels). Here, the (TTTAT)3-5 microsatellite polymorphism within intron 6 of the LDHA gene in pigeons was analysed using the HRM method. Individuals (123 homing pigeons) were genotyped using conventional PCR. Birds were classified into groups based on genotype type and the results were tested by qPCR-HRM and verified using sequencing. Based on the evaluated protocol, five genotypes were identified that vary in the number of TTTAT repeat units (3/3, 4/4, 3/4, 4/5, and 5/5). Sequencing have confirmed the results obtained with qPCR-HRM and verified that HRM is a suitable method for identification of three-allele microsatellite polymorphisms. It can be concluded that the high-resolution melting (HRM) method can be effectively used for rapid (one-step) discrimination of the (TTTAT)3-5 microsatellite polymorphism in the pigeon's LDHA gene.
format	article
author	Magdalena Jedrzejczak-Silicka Adam Lepczynski Filip Gołębiowski Daniel Dolata Andrzej Dybus
author_facet	Magdalena Jedrzejczak-Silicka Adam Lepczynski Filip Gołębiowski Daniel Dolata Andrzej Dybus
author_sort	Magdalena Jedrzejczak-Silicka
title	Application of PCR-HRM method for microsatellite polymorphism genotyping in the LDHA gene of pigeons (Columba livia).
title_short	Application of PCR-HRM method for microsatellite polymorphism genotyping in the LDHA gene of pigeons (Columba livia).
title_full	Application of PCR-HRM method for microsatellite polymorphism genotyping in the LDHA gene of pigeons (Columba livia).
title_fullStr	Application of PCR-HRM method for microsatellite polymorphism genotyping in the LDHA gene of pigeons (Columba livia).
title_full_unstemmed	Application of PCR-HRM method for microsatellite polymorphism genotyping in the LDHA gene of pigeons (Columba livia).
title_sort	application of pcr-hrm method for microsatellite polymorphism genotyping in the ldha gene of pigeons (columba livia).
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/4587776b885e4fddb81d15611b2eb08f
work_keys_str_mv	AT magdalenajedrzejczaksilicka applicationofpcrhrmmethodformicrosatellitepolymorphismgenotypingintheldhageneofpigeonscolumbalivia AT adamlepczynski applicationofpcrhrmmethodformicrosatellitepolymorphismgenotypingintheldhageneofpigeonscolumbalivia AT filipgołebiowski applicationofpcrhrmmethodformicrosatellitepolymorphismgenotypingintheldhageneofpigeonscolumbalivia AT danieldolata applicationofpcrhrmmethodformicrosatellitepolymorphismgenotypingintheldhageneofpigeonscolumbalivia AT andrzejdybus applicationofpcrhrmmethodformicrosatellitepolymorphismgenotypingintheldhageneofpigeonscolumbalivia
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Application of PCR-HRM method for microsatellite polymorphism genotyping in the LDHA gene of pigeons (Columba livia).

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