Abstract
DNA can be concatenated by hybridization of DNA fragments with protruding single-stranded termini. DNA cleavage occurring at a nucleotide containing a DNA base analogue is a useful method to obtain DNA with designed protruding termini. Here, we report a novel non-enzymatic DNA cleavage reaction for DNA concatenation. We found that DNA is cleaved at a nucleotide containing 5-ethynyluracil in a methylamine aqueous solution to generate 5'-phosphorylated DNA fragment as a cleavage product. We demonstrated that the reaction can be applied to DNA concatenation of PCR-amplified DNA fragments. This novel non-enzymatic DNA cleavage reaction is a simple practical approach for DNA concatenation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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DNA Cleavage
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DNA, Concatenated / chemistry*
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Uracil / analogs & derivatives*
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Uracil / chemistry
Substances
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DNA, Concatenated
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eniluracil
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Uracil
Grants and funding
This work was supported by the Program for Innovative Cell Biology by Innovative Technology from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan (to HRU), a Grant-in-Aid for Scientific Research (S) (Grant No. 25221004) (to HRU), a Grant-in-Aid for Scientific Research on Innovative Areas (Grant No. 23115006) from MEXT/Japan Society for the Promotion of Science (JSPS) (to HRU), a grant for the strategic programs for R&D (President's discretionary fund) of RIKEN (to HRU), an intramural Grant-in-Aid from the RIKEN Center for Developmental Biology and RIKEN Quantitative Biology Center (to HRU), a grant from Core Research for Evolutional Science and Technology (CREST) (to HRU), the RIKEN Special Postdoctoral Research Program (to EAS), a Grant-in-Aid from Japan Foundation for Applied Enzymology (to EAS), and a Grant-in-Aid for JSPS Fellows A2559890 (to YS). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.