Single‐Sperm Typing

Abstract

This unit presents protocols for sperm isolation using two different methods, amplification of simple sequence-length polymorphisms (SSLP) and/or single nucleotide polymorphisms (SNP) from single cells or whole genome-amplified single cells using primer extension preamplification (PEP), and discusses the statistical analysis of sperm-typing recombination data. Newer methods for studying recombination over very short distances (a few kilobases) using total sperm DNA and allele-specific PCR are also discussed.

Mammalian Meiotic Recombination Hot Spots

“…Single-sperm typing strategies can provide a wide sweep for active hot spots along a chromosome. However, only intervals where the totality of all hot spot activity has a discernible impact on recombination intensity will be identified since high accuracy with current single-sperm methods is laborious and expensive [for details see (4,20,42,100)]. …”

Hot and Cold Spots of Recombination in the Human Genome: the Reason We Should Find Them and How This Can Be Achieved

“…Significantly more than two loci (in one case, 17 and, in the other, 32) have been amplified from a single sperm (Lien et al 2000;Pramanik and Li 2002). The single-sperm typing method, including technical details and possible errors (Li et al 1991;Cullen and Carrington 2001;Lien et al 2002), has been reviewed elsewhere.…”