A new approach for point mutation detection based on a ligase chain reaction. |
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Authors: | A V Demchinskaya I A Shilov A S Karyagina V G Lunin O V Sergienko O L Voronina M Leiser L Plobner |
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Institution: | Institute of Agricultural Biotechnology, Timiryazevskaya ul. 42, 127550, Moscow, Russia. |
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Abstract: | A new method for the identification of point mutations is proposed. The method is based on ligase chain reaction (LCR) and it includes a procedure for correction of ligation by Cleavase. Reaction products are detected by a colorimetric method after adsorption of the resulting DNA duplexes to the solid phase. One strand of LCR products carries biotin to be bound on a streptavidin-coated microwell. Another strand contains a single-stranded region that is to be coupled with an oligonucleotide carrying a substrate for colorimetric detection. The suggested method has two advantages: (i) use of Cleavase increases the accuracy of ligation and (ii) a template independent ligation does not occur in LCR due to a special design of primers. |
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