The ligation amplification reaction (LAR)—Amplification of specific DNA sequences using sequential rounds of template-dependent ligation

A novel DNA sequence detection method that utilizes the ligation of oligonucleotide pairs that are complementary to adjacent sites on appropriate DNA templates is described. The product is increased by either linear or exponential amplification using sequential rounds of template-dependent ligation....

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Veröffentlicht in:Genomics (San Diego, Calif.) Calif.), 1989-05, Vol.4 (4), p.560-569
Hauptverfasser: Wu, Dan Y., Wallace, R.Bruce
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Sprache:eng
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Zusammenfassung:A novel DNA sequence detection method that utilizes the ligation of oligonucleotide pairs that are complementary to adjacent sites on appropriate DNA templates is described. The product is increased by either linear or exponential amplification using sequential rounds of template-dependent ligation. In the case of linear amplification, a single pair of oligonucleotides is ligated, the reaction is heated to dissociated the ligation product, and an additional round of ligation is performed. After n rounds there is a (1 + x) × n-fold amplification of product, where x is the efficiency of the ligation reaction. Exponential amplification utilizes two pairs of oligonucleotides, one complementary to the upper strand and one to the lower strand of a target sequence. The products of the ligation reaction serve as templates for subsequent rounds of ligation. In this case there is (1 + x) (n−1) -fold amplification of product after n rounds. A single base-pair mismatch between the annealed oligonucleotides and the template prevents ligation, thus allowing the distinction of single base-pair differences between DNA templates. At high template concentrations, the ligation reaction has an efficiency approaching 100%. In this report, we demonstrate the use of the ligation amplification reaction (LAR) to distinguish the normal from the sickle cell allele of the human β-globin gene. We also report the use of LAR as a detection system for polymerase chain reaction-enriched DNA sequences.
ISSN:0888-7543
1089-8646
DOI:10.1016/0888-7543(89)90280-2