Development of typing methods based on pyrosequencing technology for the analysis of six bovine genes related to marbling

RIPOLI M V; ROGBERG MUÑOZ A; LIRON J P; GIOVAMBATTISTA G

Basic and Applied Genetics

SOCIEDAD ARGENTINA DE GENÉTICA

Lugar: Buenos Aires; Año: 2013 vol. 15 p. 46 - 54

1666-0390

Several methods (PCR-RFLP, OLA, DNA sequencing, PCR SSCP and ARMS-PCR) have been developed to detect allelic variation at substitutions K232A of DGAT1, GH6.1 of GH, F279Y of GHR, R4C of LEP, I74V of FABP4, and at the transition in the thyroglobulin 5´ leader sequence in two bovine breeds. All these mutations were associated with marbling. Most of the methodologies mentioned above are manual processes and therefore increases the time spent on the assay and limit the number of analyzed animals. We describe the development of pyrosequencing-based methods applicable to the mentioned mutations. These methods were validated analyzing control DNA samples previously typed by PCR-SSCP, PCR-RFLP and DNA sequencing belonging to Aberdeen Angus and Hereford breeds. These results demonstrated that this sequence-based method would be useful in high throughput genotyping SNPs of candidate genes in bovine breeds, making this method interesting as a rapid screening method for association or population studies.