SSNIP-seq: A simple and rapid method for isolation of single-sperm nucleic acid for highthroughput sequencing
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(Published version)
Date
2022
Authors
Novakovic, S.
Tsui, V.
Semple, T.
Martelotto, L.
McCarthy, D.J.
Crismani, W.
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Drevet, J.R.
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Journal article
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PLoS ONE, 2022; 17(9):1-12
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Stevan Novakovic, Vanessa Tsui, Tim Semple, Luciano Martelotto, Davis J. McCarthy, Wayne Crismani
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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.
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© 2022 Novakovic et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.