tailieunhanh - Báo cáo thú y: " DNA methylation patterns detected by the Amplified Methylation Polymorphism Polymerase Chain Reaction (AMP PCR) technique among various cell types of bulls"

Tuyển tập các báo cáo nghiên cứu về bệnh học thý y được đăng trên tạp chí Acta Veterinaria Scandinavica cung cấp cho các bạn kiến thức về bệnh thú yđề tài: DNA methylation patterns detected by the Amplified Methylation Polymorphism Polymerase Chain Reaction (AMP PCR) technique among various cell types of bulls. | Phutikanit et al. Acta Veterinaria Scandinavica 2010 52 18 http content 52 1 18 AVS ACTAVETERINARIA SCANDINAVICA RESEARCH Open Access Different DNA methylation patterns detected by the Amplified Methylation Polymorphism Polymerase Chain Reaction AMP PCR technique among various cell types of bulls Nawapen Phutikanit1 Junpen Suwimonteerabutr1 Dion Harrison2 Michael D Occhio3 Bernie Carroll2 Mongkol Techakumphu1 Abstract Background The purpose of this study was to apply an arbitrarily primed methylation sensitive polymerase chain reaction PCR assay called Amplified Methylation Polymorphism Polymerase Chain Reaction AMP PCR to investigate the methylation profiles of somatic and germ cells obtained from Holstein bulls. Methods Genomic DNA was extracted from sperm leukocytes and fibroblasts obtained from three bulls and digested with a methylation sensitive endonuclease HpaII . The native genomic and enzyme treated DNA samples were used as templates in an arbitrarily primed-PCR assay with 30 sets of single short oligonucleotide primer. The PCR products were separated on silver stained denaturing polyacrylamide gels. Three types of PCR markers digestion resistant- digestion sensitive- and digestion dependent markers were analyzed based on the presence absence polymorphism of the markers between the two templates. Results Approximately 1 000 PCR markers per sample were produced from 27 sets of primer and most of them 90 were digestion resistant markers. The highest percentage of digestion resistant markers was found in leukocytic DNA and the lowest in fibroblastic DNA P . Spermatozoa contained a higher number of digestion sensitive markers when compared with the others vs. and in leukocytes and fibroblasts respectively P . Conclusions The powerfulness of the AMP PCR assay was the generation of methylation-associated markers without any prior knowledge of the genomic sequence. The data obtained from different primers .