Nucleic Acids: Hybridisation

Abstract

Abstract Nucleic acid hybridisation is the pairing of complementary deoxyribonucleic acid (DNA) strands to produce DNA–DNA hybrids or the pairing of complementary DNA–RNA strands to produce DNA–RNA hybrids. Nucleic acid hybridisation is the basis of many research and diagnostic applications with all relying on simple base pairing specificity of nucleic acids to generate a signal through a labelled probe. This fundamental principle has led to DNA/RNA detection and quantification on solid phase blots, DNA/RNA cytogenetic localisation on cells, detection and purification of specific DNA and comparative gene expression analysis. Most recently, principles of nucleic acid hybridisation have been combined with next generation sequencing technology to create powerful new platforms for analysis which will increase in utility as we enter this new age of genomics and personalised medicine. The concepts and applications of nucleic acid hybridisation will be discussed in this review. Key Concepts: Nucleic acid hybridisation using base paring complementary allows detection of genes, mutations and RNA permitting accurate diagnosis of disease in the clinic and providing researchers with many powerful tools to analyse and interpret their work. Traditional hybridisation technologies have enabled detection of target DNA or RNA sequences on a solid‐base membrane or on chromosomes. Development of radioactive and nonradioactive probe labelling systems allowed for significantly improved sensitivity and specificity of detection of the target nucleic acids. Nucleic acid hybridisation in quantitative analysis permits detection of mutations such as deletion, insertion and copy number variation for disease diagnosis. Incorporation of next‐generation sequencing with nucleic acid hybridisation has opened a new genomic era and has enabled high‐throughput sophisticated analysis for personalised medicine by discovering novel genes and single nucleotide polymorphisms (SNP).