Strataprep plasmid miniprep kit

The total RNA was isolated from the collected tissues and converted to cDNA using ProtoScript^®II First Strand cDNA Synthesis kit (New England Biolabs, Massachusetts, USA) and oligo (dT)₁₈ primer. Full-length cDNA sequences for SmMKS2s were obtained by polymerase chain reaction (PCR) using 1 μL of the resulting cDNA and gene-specific primers (Supplementary File 8). The PCR products were purified using Expin™ Gel SV (GeneAll Biotechnology Co., Ltd., Seoul, Korea) and cloned into pGEM-T Easy cloning vector (Promega, Wisconsin, USA). The recombinant plasmids were transformed into E. coli TOP10 competent cells. Positive clones were selected by blue/white colony screening and PCR colony. Recombinant plasmids were extracted using StrataPrep Plasmid Miniprep Kit (Agilent Technologies, California, USA) and their inserts were sequenced. Three SmMKS2 sequences have been deposited in GenBank with accession number MK990608, MK990609, and MK990610, respectively. Phylogenetic and molecular evolutionary analyses were conducted using MEGA X [22 (link)].