Arrays of complementary oligonucleotides for analysing the hybridisation behaviour of nucleic acids

• Published in: Nucleic acids research Vol. 22; no. 8; pp. 1368 - 1373
• Main Authors: Southern, E.M., Case-Green, S.C., EIder, J.K., Johnson, M., Mir, K.U., Wang, L., Williams, J.C.
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 25.04.1994
• Subjects: Analytical, structural and metabolic biochemistry; Base Sequence; Biological and medical sciences; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Glass; Molecular Sequence Data; Nucleic Acid Hybridization - instrumentation; Nucleic Acid Hybridization - methods; Nucleic acids; Oligonucleotides - chemistry; Purines; Pyrimidines; RNA - chemistry; Software; Molecular hybridization; Qualitative analysis; Oligonucleotide; Investigation method; Nucleic acid; Quantitative analysis
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/22.8.1368

Arrays of oligonucleotides corresponding to a full set of complements of a known sequence can be made in a single series of base couplings in which each base in the complement is added in turn. Coupling is carried out on the surface of a solid support such as a glass plate, using a device which applies reagents in a defined area. The device is displaced by a fixed movement after each coupling reaction so that consecutive couplings overlap only a portion of previous ones. The shape and size of the device and the amount by which it is displaced at each step determines the length of the oligonucleotides. Certain shapes create arrays of oligonucleotides from mononucleotides up to a given length in a single series of couplings. The array is used in a hybridisation reaction to a labelled target sequence, and shows the hybridisation behaviour of every oligonucleotlde in the target sequence with its complement in the array. Applications include sequence comparison to test for mutation, analysis of secondary structure, and optimisation of PCR primer and antisense ollgonucleotide design.