Analytical Sciences

Abstract − Analytical Sciences, 34(5), 627 (2018).

Improved Boronate Affinity Electrophoresis by Optimization of the Running Buffer for a Single-step Separation of piRNA from Mouse Testis Total RNA
Yusuke SATO, Daijiro IWASAWA, Kuo Ping HUI, Rena NAKAGOMI, and Seiichi NISHIZAWA
Department of Chemistry, Graduate School of Science, Tohoku University, Aoba, Sendai 980-8578, Japan
Here we examined optimization of the running buffer in boronate affinity electrophoresis for improved separation of PIWI-interacting RNA (piRNA) with 2′-O-methylated ribose in 3′-terminal nucleotide. The use of Good’s buffer, such as HEPES, significantly increased the separation efficiency for piRNA over normal RNA with free 3′-terminal ribose, and retained an ability to resolve the difference by at least 4-nucleotide lengths in the target piRNAs. We also demonstrated a single-step separation of piRNA from mouse testis total RNA.