Kazusa Nishimura, Hiroyuki Kokaji, Ko Motoki, Akira Yamazaki, Kyoka Nagasaka, Takashi Mori, Rihito Takisawa, Yasuo Yasui, Takashi Kawai, Koichiro Ushijima, Masanori Yamasaki, Hiroki Saito, Ryohei Nakano, Tetsuya Nakazaki
Next-generation sequencing (NGS) library construction often involves using restriction enzymes to decrease genome complexity, enabling versatile polymorphism detection in plants. However, plant leaves frequently contain impurities, such as polyphenols, necessitating DNA purification before enzymatic reactions. To overcome this problem, we developed a PCR-based method for expeditious NGS library preparation, offering flexibility in number of detected polymorphisms. By substituting a segment of the simple sequence repeat sequence in the MIG-seq primer set (MIG-seq being a PCR method enabling library construction with low-quality DNA) with degenerate oligonucleotides, we introduced variability in detectable polymorphisms across various crops...
March 8, 2024: Plant Journal