Simple workflow for genome and methylation analyses of ejaculated bovine spermatozoa with low sperm input
| Autor: | Sandeep K. Rajput, Bradford W Daigneault, George W. Smith |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Infertility
Male epigenome Computational biology Biology Benchmark Genome sperm Polymerase Chain Reaction General Biochemistry Genetics and Molecular Biology 03 medical and health sciences 0302 clinical medicine medicine bisulfite Animals 030304 developmental biology Epigenomics fertility 0303 health sciences 030219 obstetrics & reproductive medicine Epigenome Methylation DNA sequencing Oligospermia Sequence Analysis DNA DNA Methylation medicine.disease forensic Sperm Spermatozoa Semen Analysis genomic DNA Cattle methylation Biotechnology |
| Zdroj: | Biotechniques |
| ISSN: | 1940-9818 0736-6205 |
| Popis: | 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. METHOD SUMMARY We developed a cost-effective, user-friendly and reliable protocol (see Supplementary protocol) for DNA extraction from low-input samples of frozen-thawed bovine sperm, with a simple workflow for PCR amplification, bisulfite conversion and methylation analyses of individual amplicons. |
| Databáze: | OpenAIRE |
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