Quick two-step RNA ligation employing periodate oxidation.
The introduction of modified or labeled nucleotides into RNA is a powerful RNA engineering tool as it enables us to investigate how native RNA modifications affect RNA function and structure. It also helps in the structural analysis of RNA. A modified nucleotide can be introduced into a specific position of RNA by the method of two-step enzymatic ligation of RNA fragments. However, this method requires a complicated purification step between the two ligation steps that results in low yields of the ligation product. Here we have developed a new ligation technique employing periodate oxide that eliminates this purification step. This increases the total yield of the ligation product and makes it a faster procedure.
Kurata
Shinya
S
Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba 277-8562, Japan.
Ohtsuki
Takashi
T
Suzuki
Tsutomu
T
Watanabe
Kimitsuna
K
D000926:Anticodon / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000378:metabolism*; D012313:RNA / Q000235:genetics / Q000378:metabolism*; D012356:RNA, Transfer, Leu / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014529:Uridine / Q000235:genetics / Q000378:metabolism
D000926:Anticodon; D012356:RNA, Transfer, Leu; D010504:Periodic Acid; D012313:RNA; C009288:metaperiodate; D014529:Uridine