Number of the records: 1
Evaluation of two approaches to genotyping major histocompatibility complex class I in a passerine—CE-SSCP and 454 pyrosequencing
- 1.
SYSNO ASEP 0365694 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Evaluation of two approaches to genotyping major histocompatibility complex class I in a passerine—CE-SSCP and 454 pyrosequencing Author(s) Promerová, Marta (UBO-W)
Babik, W. (PL)
Bryja, Josef (UBO-W) RID, SAI, ORCID
Albrecht, Tomáš (UBO-W) RID, SAI, ORCID
Stuglik, M. (PL)
Radwan, J. (PL)Number of authors 6 Source Title Molecular Ecology Resources - ISSN 1755-098X
Roč. 12, č. 2 (2012), s. 285-292Number of pages 8 s. Language eng - English Country GB - United Kingdom Keywords avian ; Carpodacus erythrinus ; major histocompatibility complex ; next-generation sequencing ; scarlet rosefinch Subject RIV EG - Zoology R&D Projects IAA600930608 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) GA206/06/0851 GA ČR - Czech Science Foundation (CSF) GAP505/10/1871 GA ČR - Czech Science Foundation (CSF) CEZ AV0Z60930519 - UBO-W (2005-2011) UT WOS 000299930300011 EID SCOPUS 84856736609 DOI 10.1111/j.1755-0998.2011.03082.x Annotation MHC is linked to individual fitness and is important in evolutionary ecology and conservation genetics. Gene duplication and positive selection usually lead to high polymorphism in the MHC region, making genotyping of MHC a challenging task. We compare the performance of two methods for MHC class I genotyping in a passerine with highly duplicated MHC class I genes: CE-SSCP analysis and 454 GS FLX Titanium pyrosequencing. According to our findings, the number of MHC alleles detected by CE-SSCP is significantly lower than detected by 454. To resolve discrepancies between the two methods, we cloned and Sanger sequenced a MHC class I amplicon for an individual with high number of alleles. We found perfect congruence between cloning/Sanger sequencing and 454. Thus, in case of multi-locus amplification, CE-SSCP considerably underestimates individual MHC diversity. Numbers of alleles detected by both methods are significantly correlated, although the correlation is weak (r=0.32). Workplace Institute of Vertebrate Biology Contact Hana Slabáková, slabakova@ivb.cz, Tel.: 543 422 524 Year of Publishing 2013
Number of the records: 1