Combination of S1 nuclease and PNA for site-selective hydrolysis of double-stranded DNA. Comparison with the site-selective hydrolysis using Ce(IV)/EDTA.
The potential of the combination of SI nuclease and pseudo-complementary PNA (pcPNA) for site-selective scission of double-stranded DNA has been investigated. Through strand invasion of two pcPNAs, single-stranded portions were formed in both strands of substrate DNA. In the initial stage of the enzymatic digestion, two scission fragments were obtained due to the hydrolysis at these two gap-like sites. On prolonged reactions, however, these products (as well as the substrate DNA) were further digested to smaller fragments. Under the conditions employed here, only Ce(IV)/EDTA is available for the preparation of desired fragments from double-stranded DNA.
Yamamoto
Yoji
Y
Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Komiyama
Makoto
M
D002563:Cerium / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D004492:Edetic Acid / Q000378:metabolism*; D006868:Hydrolysis; D020135:Peptide Nucleic Acids / Q000378:metabolism*; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism*
D020135:Peptide Nucleic Acids; D002563:Cerium; D004247:DNA; D004492:Edetic Acid; D015719:Single-Strand Specific DNA and RNA Endonucleases