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HIGH-THROUGHPUT METHOD FOR DETERMINATION OF APOLIPOPROTEIN-E GENOTYPES WITH USE OF RESTRICTION DIGESTION ANALYSIS BY MICROPLATE ARRAY DIAGONAL GEL-ELECTROPHORESIS

BOLLA, MK; HADDAD, L; HUMPHRIES, SE; WINDER, AF; DAY, INM; (1995) HIGH-THROUGHPUT METHOD FOR DETERMINATION OF APOLIPOPROTEIN-E GENOTYPES WITH USE OF RESTRICTION DIGESTION ANALYSIS BY MICROPLATE ARRAY DIAGONAL GEL-ELECTROPHORESIS. CLIN CHEM , 41 (11) 1599 - 1604.

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Abstract

Molecular epidemiological research has identified the association of a common apolipoprotein E (ape E) isoform (E4 as opposed to E3), with risk both of coronary artery disease and of Alzheimer dementia. In addition, the role of apo E genotype (usually E2/E2) in Type III hyperlipidemia is well known. However, both for diagnostic and research purposes, apo E genotyping is cumbersome. The preferred approach is electrophoretic sizing of restriction digestion fragments, enabling simultaneous analysis of the two codons (112 and 158) that represent the six common genotypes (E2/E2; E2/E3; E2/E4; E3/E3; E3/E4; E4/E4). However, the consequent demands of high-yield PCR, high-resolution, high-throughput electrophoresis, and sufficient detection sensitivity have left shortfalls in published protocols. In conjunction with a high-throughput electrophoresis system we described recently, microplate array diagonal gel electrophoresis (MADGE), we have constructed extensively optimized, simplified protocols for DNA isolation from mouthwash samples for PCR setup and high-yield PCR, for restriction digestion, and for subsequent MADGE gel image analysis. The integral system enables one worker to readily undertake apo E genotyping of as many as hundreds of DNA samples per day, without special equipment.

Type:Article
Title:HIGH-THROUGHPUT METHOD FOR DETERMINATION OF APOLIPOPROTEIN-E GENOTYPES WITH USE OF RESTRICTION DIGESTION ANALYSIS BY MICROPLATE ARRAY DIAGONAL GEL-ELECTROPHORESIS
Keywords:POLYMERASE CHAIN REACTION, CORONARY ARTERY DISEASE, ALZHEIMER DEMENTIA, GEL IMAGE ANALYSIS, A-IV, DISEASE, POLYMORPHISMS, BINDING, GENE, DNA, AMPLIFICATION, POPULATIONS, FREQUENCY, DIAGNOSIS
UCL classification:UCL > School of Life and Medical Sciences > Faculty of Population Health Sciences > Institute of Cardiovascular Science

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