Open AccessSimple workflow for genome and methylation analyses of ejaculated bovine spermatozoa with low sperm input
Author(s) -
Bradford W. Daigneault,
Sandeep Rajput,
George W. Smith
Publication year - 2020
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2019-0121
Subject(s) - dna methylation , biology , epigenomics , sperm , genetics , reproductive technology , computational biology , andrology , embryo , gene , medicine , cryopreservation , gene expression
We developed a simplified workflow of gDNA extraction from ejaculated bovine sperm using a low total number of sperm and a short time frame that yields high-quality DNA suitable for downstream methylation and genome analyses. These techniques have broad implications in human biomedical sciences and agriculture, including clinical diagnoses of infertility, the identification of single-nucleotide polymorphisms and aberrant methylation patterns that can impact fertility, lower embryo development and contribute to heritable disease. The methods described here provide a reliable, simplistic approach for analyzing both the genomic and epigenomic status of whole sperm ejaculates that can be adapted for laboratory diagnostics, clinical reproductive practice and basic research.