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Methods of Sperm DNA Extraction for Genetic and Epigenetic Studies

  • Jeanine Griffin
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 927)

Abstract

High quality DNA extractions developed for mammalian somatic cells are ineffective for sperm, due mainly to the high degree of nuclear compaction in sperm. The highly specialized nuclear proteins in sperm create a chromatin structure that is at least six times denser than histone bound DNA. Unlike somatic cells, sperm DNA is highly compacted by the replacement of histones with sperm-specific low molecular weight proteins called protamines. Both the protamines and the disulfide bridges formed within and between protamines inhibit the extraction of sperm DNA by standard techniques used for somatic cells. Here we describe the guanidine thiocyanate method reported by Hossain with additional modifications resulting in high molecular weight DNA of high quality with an A260/280 ratio ranging between 1.8 and 2.0 and an A260/230 ratio of 2.0 and greater. The DNA is efficiently digested with restriction enzymes and amplified by PCR.

Key words

Sperm Protamines Guanidinium thiocyanate DNA Chaotropic agent Dithiothreitol Proteinase K 

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  1. 1.Andrology and IVF Laboratories, Division of Urology, Department of SurgeryUniversity of Utah School of MedicineSalt Lake CityUSA

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