E. A. SHADENOVA*, A. A. MAMIROVA, M. T. SEMBEKOV AND A. A. IMANBAYEVA
Institute of General Genetics and Cytology, Almaty, Kazakhstan
(e-mail : shadenova.e.a@mail.ru; Phone : (3272) 498217)
(Received : May 17, 2019; Accepted : July 20, 2019)
ABSTRACT
This article describes a method for the rapid and convenient isolation of genomic DNA from leaves of woody plants. In this article, a standard method of isolating genomic DNA from 45 rare and endemic plant species was tested. Three methods of DNA analysis were used : visual (presence or absence of white precipitate, contamination), electrophoresis and spectrophotometry. Based on the results of electrophoregrams and spectrophotometry, a decision was made to modify the isolation protocol. Modified method of DNA extraction was based on the absence of liquid nitrogen, mercaptoethanol and RNAse A. Only T1 was used. Due to the fact that the studied species contained sufficiently large amount of tannins, an increase of incubation and centrifugation time had a positive effect on the result of DNA isolation. This method allowed to obtain high quality preparations with a DNA average concentration of 140.32 µg. The resulting DNA product can be used in plants molecular genetics identification and certification studies.
Key words :DNA, leaves, extraction, spectrophotometry, electrophoresis