Methylase-limited partial NotI cleavage for physical mapping of genomic DNA.

Abstract:

UNLABELLED:Partial cleavage of DNA with the restriction endonuclease NotI (5'...GC/GGCCGC...3') is an important technique for genomic mapping. However, partial genomic cleavage with this enzyme is impaired by the agarose matrix in which the DNA must be suspended. To solve this problem we have purified the blocking methylase M. BspRI (5'...GGmCC...3') for competition digests with NotI. The resulting methylase-limited partial DNA cleavage is shown to be superior to standard techniques on bacterial genomic DNA. ABBREVIATIONS:bp, base-pair; kb, one thousand base-pairs; Mb, one million base-pairs; Tris, Tris(hydroxy-methyl)aminomethane; EDTA, (ethylenedinitrilo)tetraacetic; beta-ME, beta-mercaptoethanol; PMSF, phenyl methyl-sulfonyl fluoride; PEG, polyethyleneglycol (MW = 8000); 3H, tritium; SAM, S-adenosylmethionine; KGB, potassium glutamate buffer; DTT, dithiothreitol; IPTG, isopropyl-beta-D-thiogalactopyranoside; BSA, bovine serum albumin.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Hanish J,McClelland M

doi

10.1093/nar/18.11.3287

subject

Has Abstract

pub_date

1990-06-11 00:00:00

pages

3287-91

issue

11

eissn

0305-1048

issn

1362-4962

journal_volume

18

pub_type

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