A rapid method to identify cosmids containing rare restriction site

Xavier P. Estivill, R. Williamson

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

A procedure for identifying specific cosmid clones containing recognition sites for "rare cutting" restriction enzymes has been developed. Cosmid clones containing human inserts were selected by hybridisation to human repetitive DNA. An oligonucleotide corresponding to the NotI recognition site, eight bases long, was labelled and used to probe DNA samples from one hundred cosmids. By optimising the difference in melting characteristics between eight-base perfect match and six-base match/two base mismatch hybrids, we were able to detect the cosmids containing either NotI (8 bp match) or XmaIII/EagI (6 bp match) sites. The generation of a map for rare cutter sites along a human chromosome, or a chromosome region, should be simplified using this approach, which will enable the identification of a set of "milestones" at intervals of several hundred kilobases (kb) along the DNA sequence.

Original languageEnglish
Pages (from-to)1415-1425
Number of pages11
JournalNucleic Acids Research
Volume15
Issue number4
DOIs
Publication statusPublished - 25 Feb 1987
Externally publishedYes

Fingerprint

Cosmids
Clone Cells
DNA Probes
Human Chromosomes
Oligonucleotides
Freezing
Chromosomes
DNA
Enzymes

ASJC Scopus subject areas

  • Genetics

Cite this

A rapid method to identify cosmids containing rare restriction site. / Estivill, Xavier P.; Williamson, R.

In: Nucleic Acids Research, Vol. 15, No. 4, 25.02.1987, p. 1415-1425.

Research output: Contribution to journalArticle

@article{c35730ef8bcf4be6ba8edac4dc081282,
title = "A rapid method to identify cosmids containing rare restriction site",
abstract = "A procedure for identifying specific cosmid clones containing recognition sites for {"}rare cutting{"} restriction enzymes has been developed. Cosmid clones containing human inserts were selected by hybridisation to human repetitive DNA. An oligonucleotide corresponding to the NotI recognition site, eight bases long, was labelled and used to probe DNA samples from one hundred cosmids. By optimising the difference in melting characteristics between eight-base perfect match and six-base match/two base mismatch hybrids, we were able to detect the cosmids containing either NotI (8 bp match) or XmaIII/EagI (6 bp match) sites. The generation of a map for rare cutter sites along a human chromosome, or a chromosome region, should be simplified using this approach, which will enable the identification of a set of {"}milestones{"} at intervals of several hundred kilobases (kb) along the DNA sequence.",
author = "Estivill, {Xavier P.} and R. Williamson",
year = "1987",
month = "2",
day = "25",
doi = "10.1093/nar/15.4.1415",
language = "English",
volume = "15",
pages = "1415--1425",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "4",

}

TY - JOUR

T1 - A rapid method to identify cosmids containing rare restriction site

AU - Estivill, Xavier P.

AU - Williamson, R.

PY - 1987/2/25

Y1 - 1987/2/25

N2 - A procedure for identifying specific cosmid clones containing recognition sites for "rare cutting" restriction enzymes has been developed. Cosmid clones containing human inserts were selected by hybridisation to human repetitive DNA. An oligonucleotide corresponding to the NotI recognition site, eight bases long, was labelled and used to probe DNA samples from one hundred cosmids. By optimising the difference in melting characteristics between eight-base perfect match and six-base match/two base mismatch hybrids, we were able to detect the cosmids containing either NotI (8 bp match) or XmaIII/EagI (6 bp match) sites. The generation of a map for rare cutter sites along a human chromosome, or a chromosome region, should be simplified using this approach, which will enable the identification of a set of "milestones" at intervals of several hundred kilobases (kb) along the DNA sequence.

AB - A procedure for identifying specific cosmid clones containing recognition sites for "rare cutting" restriction enzymes has been developed. Cosmid clones containing human inserts were selected by hybridisation to human repetitive DNA. An oligonucleotide corresponding to the NotI recognition site, eight bases long, was labelled and used to probe DNA samples from one hundred cosmids. By optimising the difference in melting characteristics between eight-base perfect match and six-base match/two base mismatch hybrids, we were able to detect the cosmids containing either NotI (8 bp match) or XmaIII/EagI (6 bp match) sites. The generation of a map for rare cutter sites along a human chromosome, or a chromosome region, should be simplified using this approach, which will enable the identification of a set of "milestones" at intervals of several hundred kilobases (kb) along the DNA sequence.

UR - http://www.scopus.com/inward/record.url?scp=0023664036&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023664036&partnerID=8YFLogxK

U2 - 10.1093/nar/15.4.1415

DO - 10.1093/nar/15.4.1415

M3 - Article

VL - 15

SP - 1415

EP - 1425

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 4

ER -