Genomic DNA Extraction from Sperm

Document Type : Methodology Report

Authors

1 Infertility Research Center, Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

2 Department of Immunology, Shiraz University of Medical Sciences, Medical School, Shiraz, Iran

Abstract

Unlike the somatic cells, sperm DNA is very compact due to replacement of histones with protamines. Disulfide bridges formed within and between the protamines inhibit the extraction of sperm DNA through standard techniques used for the somatic cells. Furthermore, the spermatozoa themselves are protected by a membrane which is rich in disulfide bonds, making cell lysis very difficult. Following a comprehensive literature search, we developed a protocol for DNA extraction from sperm and semen fluid. The quality of extracted DNA was checked running on agarose gel, used for bisulfite conversion and PCR amplification.

Keywords

References

  1. Oliva R. Protamines and male infertility. Hum Reprod Update 2006; 12: 417-435
  2. Cree LH, Balhorn R, and Brewer LR. Single Molecule Studies of DNA-Protamine Interactions. Protein Pept Lett 2011; 18: 802-810
  3. Bahnak BR, Wu QY, Coulomber L, Drouet L, Kerbiriou-Nabias D, et al. A simple and efficient method for isolating high molecular weight DNA from mammalian sperm. Nucleic Acids Res 1988; 16: 1208
  4. Griffin J. Methods of Sperm DNA Extraction for Genetic and Epigenetic Studies. Methods Mol Biol 2013;927:379–384
  5. Pacheco S, Houseman E, Christensen B, Marsit C, Kelsey K, et al. Integrative DNA Methylation and Gene Expression Analyses Identify DNA Packaging and Epigenetic regulatory genes associated with low Motility sperm. PLoS ONE 2011; 6 (6): e20280
  6. Hossain A. Modified guanidinium thiocyanate method for human sperm DNA isolation. Mol Hum Reprod 1997; 3: 953-956
  7. Wu H, De Gannes MK, Luchetti G, and Pilsner JR. Rapid method for the isolation of mammalian sperm DNA. Biotechniques 2015; 58:293-300
  8. Weyrich A. Preparation of genomic DNA from mammalian sperm. Curr Protoc Mol Biol 2012;Chapter 2:Unit2 13 1-3
  9. Gebert C, Wrenzycki C, Hermann D, Groger D, Reinhardt R, et al. The bovin IGF2 gene is differentially methylated in oocyte and sperm DNA. GENOMICS 2006;88: 222–229
  10. Marques C, Costa P, Vaz B, Carvalho F, Fernandes S, et al. Abnormal methylation of imprinted genes in human sperm is associated with oligozoospermia. Mol Hum Reprod 2007; 14: 67-74
Journal of Advanced Medical Sciences and Applied Technologies
Volume 1, Issue 2 - Serial Number 2
5
December 2015
Pages 120-121

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