Analysis of DNA polymorphisms on the human Y-chromosome by microchip electrophoresis

Abstract

Validation of microchip electrophoresis in DNA analysis has been carried out using an Agilent 2100 Bioanalyzer. With a DNA 500 Assay Kit, the reproducibility and accuracy of fragment sizing of a 10 bp DNA ladder have been shown to be satisfactory with the relative standard deviation and the relative error mostly below 1.0 and 5.0% (n = 12), respectively. Both intraday and interday validations of fragment sizing and quantitation have also been performed with a 7500 Assay Kit (n = 48). Although the results of quantitation are not as good as that of sizing, due to the manual introduction of samples and markers into the chip wells, they are still sufficient to carry out further analyses of practical samples. Based on such reliable results, fast analysis of DNA polymorphisms on the human Y-chromosome has been realized with microchip electrophoresis. The total analysis times of three genomic polymorphisms on the Y-chromosome, Y Alu polymorphism, 47z/StuI, and 12f2, are all within 100 s, and the relative standard deviation and relative error of fragment sizes are below 3.5 and 3.7%, respectively. In addition, a mixture of nine DNA markers on the human Y-chromosome related to examine the cause of spermatogenic failure have been separated successfully with the smallest fragment size difference of 7 bp. Our results demonstrate the potential of microchip electrophoresis in polymorphism analysis with the advantages of high speed, good reproducibility, high precision, and high resolution.

Range of Separation of Potential Tool for Bioseparation, Microchip Electrophoresis System, for DNA Polymorphisms on the Human Y-Chromosome

“…3 Indeed, the human Ychromosome has unique characteristics in genetics because it is For the requirement of a high, fast and sufficient technology to suit the needs of 21st century biotechnology, the separation range of a microchip electrophoresis system was studied. Two DNA fragments on the human Y-chromosome, SY594 (82 bp) and 12f2 (88 bp), were successfully separated with a reproducibility of 1.9% and an accuracy of 2.8%.…”

“…The Human Genome Project (HGP) is presently moving quickly to the post-genome era, which will stimulate investigations of single nucleotide polymorphism (SNP), mutation analysis and proteome analysis. 3 This will lead to gene therapy and DNA diagnosis of human diseases, which will result in a new class of drugs. 4 This significant progress is partly due to a huge leap in electrophoresis technology.…”

Rapid Multiplexing and Simultaneous Detection of Human Spermatogenetic Failure with a 12 Lane Microchip Electrophoresis System

“…Microchip electrophoresis as an ultrafast and reliable tool that can be utilized in screening assays for the diagnosis of presymptomatic disease states and detection of the underlying causes of non obstructive azoospermia and hypo spermatogenesis, i.e., spermatogenetic failure. [18][19][20] Approximately 20% of men who seek help at infertility clinics present with non obstructive oligo or azoospermia. Most of these men are chromosomally normal but may have microdeletions in the AZF regions of their Y chromosomes, which cause them to have low sperm counts or severe forms of spermatogenic failure.…”

Determination of human blood glucose levels using microchip electrophoresis

“…This approach has been successfully used to analyze DNA fragments [22][23][24][25], amino acids [26], proteins [27,28], and carbohydrates [29]. Although several microchip manufacturing techniques have been reported for the detection of blood glucose concentration, suitable analysis will require some elaboration of microchip design, detection systems, and electrodes [30][31][32][33][34][35][36].…”