Improved Boronate Affinity Electrophoresis by Optimization of the Running Buffer for a Single-step Separation of piRNA from Mouse Testis Total RNA

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...

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Published inAnalytical Sciences Vol. 34; no. 5; pp. 627 - 630
Main Authors SATO, Yusuke, IWASAWA, Daijiro, HUI, Kuo Ping, NAKAGOMI, Rena, NISHIZAWA, Seiichi
Format Journal Article
LanguageEnglish
Published Singapore The Japan Society for Analytical Chemistry 2018
Springer Nature Singapore
Japan Science and Technology Agency
Subjects
2′-O-methylatyion
Affinity
Analytical Chemistry
Boronate affinity electrophoresis
Buffers
Chemistry
Electrophoresis
Optimization
phenylboronic acid
piRNA
Ribonucleic acid
Ribose
RNA
Separation
2
methylatyion
phenylboronic acid
Boronate affinity electrophoresis
separation
piRNA
2′-O-methylatyion
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Summary: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.
Bibliography:ObjectType-Article-1
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ISSN:0910-6340
1348-2246
DOI:10.2116/analsci.17N024