Affinity Isolation of Defined Genomic Fragments Cleaved by Nuclease S1-based Artificial Restriction DNA Cutter.
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
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Genoma Humano
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Cromatografía de Afinidad
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Ácidos Nucleicos de Péptidos
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División del ADN
Idioma:
En
Revista:
Curr Protoc Nucleic Acid Chem
Año:
2019
Tipo del documento:
Article
País de afiliación:
Japón