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DNA Microarrays

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...

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Visual Detection of Multiple Nucleic Acids in a Capillary Array

Visual Detection of Multiple Nucleic Acids in a Capillary Array

Published on: November 15, 2017

Two-dye based arrayed primer extension for simultaneous multigene detection in lipid metabolism.

Nutjaree Jeenduang¹, Sureerut Porntadavity², Markus von Nickisch-Rosenegk³

¹School of Allied Health Science and Public Health, Walailak University, Nakhon Si Thammarat 80161, Thailand; Department of Nanobiotechnology and Nanomedicine, Fraunhofer Institute for Biomedical Engineering (IBMT), Potsdam-Golm 14476, Germany.

Clinica Chimica Acta; International Journal of Clinical Chemistry

|January 17, 2015

Summary

A new microarray assay enables rapid, simultaneous detection of multiple genetic risk factors for cardiovascular disease (CVD), including LDLR, APOB, PCSK9, CETP, and APOE mutations and polymorphisms.

Keywords:

Arrayed primer extension (APEX)Genotyping Lipid metabolism Microarray

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Area of Science:

Genetics
Molecular Biology
Biotechnology

Background:

Cardiovascular disease (CVD) is a leading global cause of mortality.
Genetic factors influencing lipid metabolism, such as LDLR, APOB, PCSK9, CETP, and APOE, are associated with CVD risk.

Purpose of the Study:

To develop a simultaneous multigene detection assay for key genetic risk factors of CVD.
To establish an efficient method for screening genetic variations linked to hypercholesterolemia.

Main Methods:

Development of a two-dye based arrayed primer extension (APEX) microarray assay.
Simultaneous detection of LDLR, APOB, PCSK9, CETP, and APOE genes.
Uniplex PCR amplification of DNA templates from 8 individual samples.

Main Results:

Optimized APEX reaction conditions determined (55°C hybridization, 50-150bp template size).
Complete assay workflow, including PCR and analysis, completed in 6 hours.
Successfully identified 48 distinct genotypes across 8 DNA samples.

Conclusions:

The APEX microarray assay provides a robust and rapid method for genetic screening.
This assay is a versatile tool for identifying hypercholesterolemia patients with specific genetic profiles.