Abstract
Genetic studies in humans have been limited by various factors, including small family size and diploidy of the human genome. The ability to use individual spermatozoa as subjects has significantly facilitated these studies. However, because each sperm usually contains only one copy of the genome and the sensitivity of the available detection methods was low, single sperm could not be used for large-scale genetic analysis. After a series of enhancements, a high-throughput genotyping system has been developed. With this system, more than 1000 genetic markers consisting of single-nucleotide polymorphisms (SNPs) in a single sperm genome can be amplified to a detectable amount in a single tube. Sequences amplified from different polymorphic loci are then resolved by hybridizing these sequences to the corresponding probes spotted on a microarray. The allelic sequences of each SNP are then discriminated by the commonly used allele-specific single-base-extension assay with dideoxyribonucleotides labeled with different fluorescent dyes. Using single sperm cells as subjects, the highly sensitive and high-throughput multiplex genotyping system was used to analyze the physical structure of the human immunoglobulin heavy chain variable region that contains highly repetitive sequences and may not be easily analyzed with conventional methods. This system was also used to understand the contribution of meiotic recombination to haplotype structure formation in the human genome. The system may have various applications including study of genetic factors underlying male infertility.
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Li, H. et al. (2007). Microarray Analysis of a Large Number of Single-Nucleotide Polymorphisms in Individual Human Spermatozoa. In: Carrell, D.T. (eds) The Genetics of Male Infertility. Humana Press. https://doi.org/10.1007/978-1-59745-176-5_4
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DOI: https://doi.org/10.1007/978-1-59745-176-5_4
Publisher Name: Humana Press
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