About: Analysis of major histocompatibility complex class II gene in water voles using capillary electrophoresis-single stranded conformation polymorphism

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About: Analysis of major histocompatibility complex class II gene in water voles using capillary electrophoresis-single stranded conformation polymorphism Goto Sponge NotDistinct Permalink

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rdf:type	skos:Concept http://linked.opendata.cz/ontology/domain/vavai/Vysledek
Description	Analysis of a single strand conformation polymorphism (SSCP) using capillary electrophoresis (CE) is a novel method to study polymorphism of DNA sequences in large scale population studies. We optimized CE-SSCP analysis to study the major histocompatibility complex (MHC) class II alpha gene (DQA) polymorphism. Short-chain linear polyacrylamide (6%) as sieving matrix, TrisCl (pH 8.5) as buffer for sample dilution, and 27 degrees C, 9 kV as electrophoresis parameters were suitable for sufficient resolution of all alleles. We found that almost 25% of clones contained a PCR (polymerase chain reaction) artefact and strict criteria have to be applied when using cloning and sequencing to analyse the allelic diversity of MHC genes. Analysis of a single strand conformation polymorphism (SSCP) using capillary electrophoresis (CE) is a novel method to study polymorphism of DNA sequences in large scale population studies. We optimized CE-SSCP analysis to study the major histocompatibility complex (MHC) class II alpha gene (DQA) polymorphism. Short-chain linear polyacrylamide (6%) as sieving matrix, TrisCl (pH 8.5) as buffer for sample dilution, and 27 degrees C, 9 kV as electrophoresis parameters were suitable for sufficient resolution of all alleles. We found that almost 25% of clones contained a PCR (polymerase chain reaction) artefact and strict criteria have to be applied when using cloning and sequencing to analyse the allelic diversity of MHC genes. (en) Kapilární elektroforéza SSCP je nová metoda studující polymorfismus DNA sekvencí zejména v populačně-genetických studiích. V práci je popisována optimalizace této metody při studiu polymorfismu genu DQA, který náleží mezi MHC geny skupiny II. Ideální podmínky kapilární elektroforézy, které umožnily odlišení téměř všech alel byly následující: SLPA jako dělící polymer, TrisCl (pH 8,5) jako pufr pro ředění vzorku a 27°C a 9 kV jako parametry elektroforézy. Zároveň jsme zjistili, že téměř 25% všech analyzovaných klonů obsahovalo PCR artefakt, proto by při analýzách MHC variability měla být uplatňována striktní kriteria pro identifikaci nových alel. (cs)
Title	Analysis of major histocompatibility complex class II gene in water voles using capillary electrophoresis-single stranded conformation polymorphism Analýza genu hlavního histokompatibilního komplexu třídy II u hryzce vodního použitím analýzy SSCP v kapiláře (cs) Analysis of major histocompatibility complex class II gene in water voles using capillary electrophoresis-single stranded conformation polymorphism (en)
skos:prefLabel	Analysis of major histocompatibility complex class II gene in water voles using capillary electrophoresis-single stranded conformation polymorphism Analýza genu hlavního histokompatibilního komplexu třídy II u hryzce vodního použitím analýzy SSCP v kapiláře (cs) Analysis of major histocompatibility complex class II gene in water voles using capillary electrophoresis-single stranded conformation polymorphism (en)
skos:notation	RIV/68081766:_____/05:00000406!RIV/2005/AV0/A67005/N
http://linked.open.../vavai/riv/strany	173;176
http://linked.open...avai/riv/aktivita	Z
http://linked.open...avai/riv/aktivity	Z(AV0Z6093917)
http://linked.open...iv/cisloPeriodika	1
http://linked.open...vai/riv/dodaniDat	2005
http://linked.open...aciTvurceVysledku	Bryja, Josef
http://linked.open.../riv/druhVysledku	J - Článek v odborném periodiku
http://linked.open...iv/duvernostUdaju	S - Úplné a pravdivé údaje nepodléhající ochraně podle zvláštních právních předpisů
http://linked.open...titaPredkladatele	Ústav biologie obratlovců AV ČR, v. v. i.
http://linked.open...dnocenehoVysledku	512070
http://linked.open...ai/riv/idVysledku	RIV/68081766:_____/05:00000406
http://linked.open...riv/jazykVysledku	eng - angličtina
http://linked.open.../riv/klicovaSlova	water vole;population genetics (en)
http://linked.open.../riv/klicoveSlovo	population genetics water vole
http://linked.open...odStatuVydavatele	GB - Spojené království Velké Británie a Severního Irska
http://linked.open...ontrolniKodProRIV	[4DA37311B11B]
http://linked.open...i/riv/nazevZdroje	Molecular ecology notes
http://linked.open...in/vavai/riv/obor	EB
http://linked.open...ichTvurcuVysledku	1 (xsd:int)
http://linked.open...cetTvurcuVysledku	4 (xsd:int)
http://linked.open...UplatneniVysledku	2005
http://linked.open...v/svazekPeriodika	5
http://linked.open...iv/tvurceVysledku	Bryja, Josef Cosson, J.-F. Charbonnel, N. Galan, M.
http://linked.open...n/vavai/riv/zamer	Diversity and life history of free-living vertebrates and implication of results in the strategy of conservation and sustainable management of natural resources
issn	1471-8278
number of pages	4 (xsd:int)

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