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

Archive ouverte

Bryja, Josef | Galan, Maxime | Charbonnel, Nathalie | Cosson, Jean-Francois, J.-F.

Edité par CCSD ; Wiley-Blackwell -

Correspondance: bryja@brno.cas.cz. International audience. 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 ° 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

• Description
• Sujet/Public
• Outil
• Outil d'anticipation
• Mémoire et thèse
• Gestion