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Nucleic Acids Research, 2003, Vol. 31, No. 22 e145
© 2003 Oxford University Press

Article

Quick two-step RNA ligation employing periodate oxidation

Shinya Kurata, Takashi Ohtsuki*, Tsutomu Suzuki and Kimitsuna Watanabe

Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba 277-8562, Japan

*To whom correspondence should be addressed. Tel/Fax: +81 471 36 3606; Email: ohtsuki{at}kwl.t.u-tokyo.ac.jp
Correspondence may also be addressed to Kimitsuna Watanabe. Tel/Fax: +81 471 36 3601; Email: kw{at}kwl.t.u-tokyo.ac.jp
The authors wish it to be known that, in their opinion, the first two authors should regarded as joint First Authors

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.


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