Open AccessThe Applicability of Three DNA Isolation Methods in SSR Analysis of Hexaploid Plum (Prunus domestica L.) Cultivars
Author(s) -
Jasna Hanjalić,
Lejla Lasić,
Fuad Gaši,
Mekjell Meland,
Jasmin Grahić,
Belma Kalamujić Stroil
Publication year - 2018
Publication title -
genetics and applications
Language(s) - English
Resource type - Journals
eISSN - 2566-431X
pISSN - 2566-2937
DOI - 10.31383/ga.vol2iss1pp1-7
Subject(s) - dna extraction , prunus , genomic dna , biology , dna , microsatellite , isolation (microbiology) , cultivar , extraction (chemistry) , botany , polymerase chain reaction , chromatography , genetics , gene , bioinformatics , chemistry , allele
The main goal of any DNA extraction procedure is to ensure reliable and reproducible results in a simple, fast and inexpensive manner. When it comes to plant tissues, this goal is challenging to achieve due to the presence of a variety of metabolites that interfere with DNA during isolation and downstream analysis. In this study, we compared the efficiency of three methods for DNA extraction from plum kernels: 1) the standard CTAB Soltis method which is the most common protocol for DNA extraction from various plant tissues (seeds, young leafs, mature leafs, root); 2) CTAB-based method originally described for DNA isolation from medicinal plants with high levels of secondary metabolites; 3) and one of various commercially available kits. The usefulness of the obtained DNA was evaluated by SSR analysis with seven microsatellite markers. Although the latter two extraction protocols retrieved genomic DNA that gave positive PCR results, only DNA isolated by kit produced full SSR profile