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PCR-based isolation of multigene families: lessons from the avian MHC class IIB
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SYSNO ASEP 0430104 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title PCR-based isolation of multigene families: lessons from the avian MHC class IIB Author(s) Burri, R. (CH)
Promerová, Marta (UBO-W)
Goebel, J. (CH)
Fumagalli, L. (CH)Number of authors 4 Source Title Molecular Ecology Resources - ISSN 1755-098X
Roč. 14, č. 4 (2014), s. 778-788Number of pages 11 s. Language eng - English Country GB - United Kingdom Keywords Birds ; Major histocompatibility complex ; Multigene families ; PCR bias Subject RIV EG - Zoology R&D Projects GAP505/10/1871 GA ČR - Czech Science Foundation (CSF) Institutional support UBO-W - RVO:68081766 UT WOS 000337611400011 EID SCOPUS 84902304355 DOI 10.1111/1755-0998.12234 Annotation The amount of sequence data available today highly facilitates the access to genes from many gene families. Primers amplifying the desired genes over a range of species are readily obtained by aligning conserved gene regions, and laborious gene isolation procedures can often be replaced by quicker PCR-based approaches. However, in the case of multigene families, PCR-based approaches bear the often ignored risk of incomplete isolation of family members. This problem is most prominent in gene families with highly variable and thus unpredictable number of gene copies among species, such as in the major histocompatibility complex (MHC). In this study, we (i) report new primers for the isolation of the MHC class IIB (MHCIIB) gene family in birds and (ii) share our experience with isolating MHCIIB genes from an unprecedented number of avian species from all over the avian phylogeny. We report important and usually underappreciated problems encountered during PCR-based multigene family isolation and provide a collection of measures to help significantly improving the chance of successfully isolating complete multigene families using PCR-based approaches. Workplace Institute of Vertebrate Biology Contact Hana Slabáková, slabakova@ivb.cz, Tel.: 543 422 524 Year of Publishing 2015
Number of the records: 1