Multiplex PCR with Multichannel Microchip Electrophoresis: An Ultrafast Analysis for Genetic Diseases

• Published in: Journal of chromatographic science Vol. 43; no. 5; pp. 221 - 225
• Main Authors: Jabasini, Mohammad, Ewis, Ashraf A., Xu, Feng, Mohamadi, Momamad R., Ping, Guichen, Shinka, Toshikatsu, Nakahori, Yutaka, Baba, Yoshinobu
• Format: Journal Article
• Language: English
• Published: Niles, IL Oxford University Press 01.05.2005; Preston Publications
• Subjects: Analytical, structural and metabolic biochemistry; Base Sequence; Biological and medical sciences; Chromosome Deletion; Chromosomes, Human, Y; DNA Primers; Dna, deoxyribonucleoproteins; Electrophoresis, Microchip; Fundamental and applied biological sciences. Psychology; Genetic Diseases, Inborn - diagnosis; Humans; Male; Nucleic acids; Oligonucleotide Array Sequence Analysis; Polymerase Chain Reaction - methods; Reproducibility of Results; Human; Fluorescence detector; Chemical analysis; Genetic marker; Multiplex polymerase chain reaction; Genomics; Reaction product; DNA chip; System on a chip; Y-Chromosome; Genetic disease; Gene amplification; Rapid technique; DNA; Microequipment; Electrophoresis; Diagnosis; Miniaturization; Microfluidics; Polymorphism
• ISSN: 0021-9665; 1945-239X
• DOI: 10.1093/chromsci/43.5.221

A Y chromosomal polymorphic markers screening strategy using a multiplex polymerase chain reaction (PCR) and DNA microchip electrophoresis technology has recently been developed. It is a part of the human Y chromosome haplotyping system for studying Japanese population genetics and its relationship with male spermatogenic failure. This strategy is based on optimizing and modifying the primer set concentrations while keeping all other components of the PCR mixtures and conditions similar to those of a singleplex PCR. Well-balanced PCR products are obtained without changing even the DNA oligomer melting temperatures. Here, a panel of primer sets are used to amplify two groups of Y chromosome markers. The first consists of five markers and the second consists of seven markers. Both are possibly deleted in infertile men. The microchip electrophoresis technology is fast and sensitive, enables direct molecular typing of several Y chromosomal markers, and is separated by a difference of as many as six base pairs.