Work

SSNIP-seq: A simple and rapid method for isolation of single-sperm nucleic acid for high-throughput sequencing

Year: 2022

Type: article

Abstract: We developed a simple and reliable method for the isolation of haploid nuclei from fresh and frozen testes. The described protocol uses readily available reagents in combination with flow cytometry to separate haploid and diploid nuclei. The protocol can be completed within 1 hour and the resulting individual haploid nuclei have intact morphology. The isolated nuclei are suitable for library preparation for high-throughput DNA and RNA sequencing using bulk or single nuclei. The protocol was optimised with mouse testes and we anticipate that it can be applied for the isolation of mature sperm from other mammals including humans.

Source: PLoS ONE

DOI: 10.1371/journal.pone.0275168

Authors: Stevan Novakovic, Vanessa Tsui, Tim Semple, Luciano Martelotto, Davis J. McCarthy, Wayne Crismani

Institutions: St Vincents Institute of Medical Research, The University of Melbourne

Language: English

Topic: Sperm and Testicular Function

Subfield: Reproductive Medicine

Field: Medicine

Domain: Health Sciences

Open Access status: gold

Estimated APC paid: $1,805

Funders: Victorian Cancer Agency, Marshfield Clinic Research Foundation, Medical Research Council, National Health and Medical Research Council