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Affinity Isolation of Defined Genomic Fragments Cleaved by Nuclease S1-based Artificial Restriction DNA Cutter.
Rajendran, Arivazhagan; Shigi, Narumi; Sumaoka, Jun; Komiyama, Makoto.
Afiliación
  • Rajendran A; Institute of Advanced Energy, Kyoto University, Kyoto, Japan.
  • Shigi N; Life Science Center of Tsukuba Advanced Research Alliance, University of Tsukuba, Ibaraki, Japan.
  • Sumaoka J; Life Science Center of Tsukuba Advanced Research Alliance, University of Tsukuba, Ibaraki, Japan.
  • Komiyama M; Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Curr Protoc Nucleic Acid Chem ; 76(1): e76, 2019 03.
Article en En | MEDLINE | ID: mdl-30753751
ABSTRACT
The human genome is highly susceptible to various modifications, lesions, and damage. To analyze lesions and proteins bound to a defined region of the human genome, the genome should be fragmented at desired sites and the region of interest should be isolated. The few available methods for isolating a desired region of the human genome have serious drawbacks and can only be applied to specific sequences or require tedious experimental procedures. We have recently developed a novel method to isolate a desired fragment of the genome released by site-specific scission of DNA using a pair of pseudo-complementary peptide nucleic acids (pcPNAs) and S1 nuclease. When conjugated to biotin, one of the pcPNAs can be used to affinity purify the cleavage product. Here we report a detailed protocol to isolate defined kilobase-length DNA fragments that can be applied to plasmid or genomic DNA and is not limited by sequence. © 2019 by John Wiley & Sons, Inc.
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Texto completo: 1 Bases de datos: MEDLINE Asunto principal: ADN de Cadena Simple / Genoma Humano / Cromatografía de Afinidad / Ácidos Nucleicos de Péptidos / División del ADN Idioma: En Revista: Curr Protoc Nucleic Acid Chem Año: 2019 Tipo del documento: Article País de afiliación: Japón

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: ADN de Cadena Simple / Genoma Humano / Cromatografía de Afinidad / Ácidos Nucleicos de Péptidos / División del ADN Idioma: En Revista: Curr Protoc Nucleic Acid Chem Año: 2019 Tipo del documento: Article País de afiliación: Japón