Abstract
Using the Diamond DNA kit, high quality nuclear DNA was isolated from dry leaves of the endemic species Scutellaria tuvensis. The selection of primers for ISSR analysis of genetic polymorphism of natural populations is described. During the experiment, 22 primers were tested, their effectiveness was assessed on a point scale. When assessing the primers, the number of reproducible amplified DNA fragments, the clarity and brightness of the obtained fragments, and only distinctive bands were taken into account. As a result, 10 ISSR markers were selected that are the most informative for assessing the population diversity of the species.
Highlights
Using the Diamond DNA kit, high quality nuclear DNA was isolated from dry leaves of the endemic species Scutellaria tuvensis
No studies have been conducted on sequencing the genome of S. tuvensis, to identify DNA polymorphism in populations of the species, PCR-based methods can be used, which do not need any prior information about the target sequences in the genome or its parts
One of the most common methods for detecting genetic polymorphism in plants that does not require knowledge of the nucleotide sequences of the genome is ISSR analysis (Inter Simple Sequence Repeats), which uses primers usually 16 −25 bp long, they consist of tandem short 2-4 nucleotide repeats and anchored at 3' or 5' end with 1 to 4 degenerate bases extended into the flanking sequences [5,6,7]
Summary
Using the Diamond DNA kit, high quality nuclear DNA was isolated from dry leaves of the endemic species Scutellaria tuvensis. No studies have been conducted on sequencing the genome of S. tuvensis, to identify DNA polymorphism in populations of the species, PCR-based methods can be used, which do not need any prior information about the target sequences in the genome or its parts. The aim of this work is to screen ISSR markers and select the most informative primers for assessing the genetic polymorphism of natural populations of S. tuvensis.
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