A novel procedure for selective cloning of Noti linking fragments from mammalian genomes

Takashi Ito, Yoshiyuki Sakaki

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

A novel procedure has been developed for selective cloning of NotI linking fragments from mammalian genomes. Since the majority of the NotI sites in mammalian genomes are considered to be localized in so-called HTF (Hpall tiny fragment) islands, an HTF library was constructed as an initial step to enrich the Notl sites. The plasmid DNAs were isolated en masse from the HTF library and digested with Notl. Linearized plasmid DNAs derived from Notl linking clones were efficiently separated from undigested circular DNAs by an unique pulsed field polyacrylamide gel electrophoresis (PF-PAGE). The linear DNAs were eluted from the gel, recircularizcd with T4 DNA ligase and introduced into E.coli cells. About 95% of the transformants were found to contain NotI linking fragments. The procedure will thus provide a simple and useful way of collecting Notl linking fragments for long range physical mapping of mammalian genomes.

Original languageEnglish
Pages (from-to)9177-9184
Number of pages8
JournalNucleic acids research
Volume16
Issue number19
DOIs
Publication statusPublished - Oct 11 1988
Externally publishedYes

Fingerprint

Organism Cloning
Genome
Libraries
DNA
Plasmids
DNA Ligases
Circular DNA
Chromosome Mapping
Pulsed Field Gel Electrophoresis
Islands
Polyacrylamide Gel Electrophoresis
Clone Cells
Gels
Escherichia coli

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

A novel procedure for selective cloning of Noti linking fragments from mammalian genomes. / Ito, Takashi; Sakaki, Yoshiyuki.

In: Nucleic acids research, Vol. 16, No. 19, 11.10.1988, p. 9177-9184.

Research output: Contribution to journalArticle

@article{853ff44b8a2845ec8b50c7c38e0c96fe,
title = "A novel procedure for selective cloning of Noti linking fragments from mammalian genomes",
abstract = "A novel procedure has been developed for selective cloning of NotI linking fragments from mammalian genomes. Since the majority of the NotI sites in mammalian genomes are considered to be localized in so-called HTF (Hpall tiny fragment) islands, an HTF library was constructed as an initial step to enrich the Notl sites. The plasmid DNAs were isolated en masse from the HTF library and digested with Notl. Linearized plasmid DNAs derived from Notl linking clones were efficiently separated from undigested circular DNAs by an unique pulsed field polyacrylamide gel electrophoresis (PF-PAGE). The linear DNAs were eluted from the gel, recircularizcd with T4 DNA ligase and introduced into E.coli cells. About 95{\%} of the transformants were found to contain NotI linking fragments. The procedure will thus provide a simple and useful way of collecting Notl linking fragments for long range physical mapping of mammalian genomes.",
author = "Takashi Ito and Yoshiyuki Sakaki",
year = "1988",
month = "10",
day = "11",
doi = "10.1093/nar/16.19.9177",
language = "English",
volume = "16",
pages = "9177--9184",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "19",

}

TY - JOUR

T1 - A novel procedure for selective cloning of Noti linking fragments from mammalian genomes

AU - Ito, Takashi

AU - Sakaki, Yoshiyuki

PY - 1988/10/11

Y1 - 1988/10/11

N2 - A novel procedure has been developed for selective cloning of NotI linking fragments from mammalian genomes. Since the majority of the NotI sites in mammalian genomes are considered to be localized in so-called HTF (Hpall tiny fragment) islands, an HTF library was constructed as an initial step to enrich the Notl sites. The plasmid DNAs were isolated en masse from the HTF library and digested with Notl. Linearized plasmid DNAs derived from Notl linking clones were efficiently separated from undigested circular DNAs by an unique pulsed field polyacrylamide gel electrophoresis (PF-PAGE). The linear DNAs were eluted from the gel, recircularizcd with T4 DNA ligase and introduced into E.coli cells. About 95% of the transformants were found to contain NotI linking fragments. The procedure will thus provide a simple and useful way of collecting Notl linking fragments for long range physical mapping of mammalian genomes.

AB - A novel procedure has been developed for selective cloning of NotI linking fragments from mammalian genomes. Since the majority of the NotI sites in mammalian genomes are considered to be localized in so-called HTF (Hpall tiny fragment) islands, an HTF library was constructed as an initial step to enrich the Notl sites. The plasmid DNAs were isolated en masse from the HTF library and digested with Notl. Linearized plasmid DNAs derived from Notl linking clones were efficiently separated from undigested circular DNAs by an unique pulsed field polyacrylamide gel electrophoresis (PF-PAGE). The linear DNAs were eluted from the gel, recircularizcd with T4 DNA ligase and introduced into E.coli cells. About 95% of the transformants were found to contain NotI linking fragments. The procedure will thus provide a simple and useful way of collecting Notl linking fragments for long range physical mapping of mammalian genomes.

UR - http://www.scopus.com/inward/record.url?scp=0023681272&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023681272&partnerID=8YFLogxK

U2 - 10.1093/nar/16.19.9177

DO - 10.1093/nar/16.19.9177

M3 - Article

C2 - 2459661

AN - SCOPUS:0023681272

VL - 16

SP - 9177

EP - 9184

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 19

ER -