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PMID: 4069995 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Reduced cleavage by sodium hydroxide of methyladenine in DNA sequencing.

Nucleic acids research ·Vol. 13 ·No. 21 ·1985-11-11 ·Pages 7639-45

Okawa N, Suyama Y, Kaji A

Abstract

When the Maxam and Gilbert DNA sequencing method which is modified by Bencini et al. (Biotechniques Jan/Feb pp4-5, 1984) is applied to DNA containing methylated adenine in a GATC sequence, the cleavage reaction by sodium hydroxide is found to be greatly reduced in comparison to that of non-methylated adenine. Thus, a faint band in A greater than C reaction suggests a methyl adenine and can be used for its detection. That the faint band corresponds to a methyladenine was confirmed by Sanger sequencing of the same fragment and further by Maxam and Gilbert sequencing of the complementary strand of DNA, which was replicated in an E. coli strain either having or lacking methylation enzymes.

MeSH Terms
Adenine/analogs & derivatives,analysis Base Sequence Chemical Phenomena Chemistry DNA/genetics Indicators and Reagents Methylation Phosphorus Radioisotopes Sodium Hydroxide
Chemicals
Indicators and Reagents Phosphorus Radioisotopes Sodium Hydroxide DNA Adenine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Okawa N
Suyama Y
Kaji A
References (17)
17 references, click to expand
  1. Thermosensitive replication of a kanamycin resistance factor.
    J Bacteriol. 1967 Sep;94(3):687-90 PMID: 4166554
  2. Identification of an Rts1 DNA fragment conferring temperature-dependent instability to vector plasmids.
    Plasmid. 1985 Mar;13(2):88-98 PMID: 2987995
  3. Isolation of deoxyribonucleic acid methylase mutants of Escherichia coli K-12.
    J Bacteriol. 1973 Jun;114(3):1143-50 PMID: 4576399
  4. Location of DNA methylation genes on the Escherichia coli K-12 genetic map.
    Mol Gen Genet. 1973 Dec 14;127(1):47-55 PMID: 4589344
  5. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  6. Detection of 5-methylcytosine in DNA sequences.
    Nucleic Acids Res. 1978 May;5(5):1479-85 PMID: 351561
  7. Recognition sequence of the dam methylase of Escherichia coli K12 and mode of cleavage of Dpn I endonuclease.
    J Biol Chem. 1979 Feb 25;254(4):1408-13 PMID: 368070
  8. Immunochemical detection of N6-methyladenine in DNA.
    Biochim Biophys Acta. 1979 Aug 29;564(1):23-30 PMID: 534639
  9. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  10. A cautionary note on the use of certain restriction endonucleases with methylated substrates.
    Gene. 1980 Oct;11(1-2):169-71 PMID: 6254852
  11. DNA methylation and its possible biological roles.
    Prog Nucleic Acid Res Mol Biol. 1981;25:33-52 PMID: 6261297
  12. Improvements of DNA sequencing gels.
    Anal Biochem. 1981 Aug;115(2):450-7 PMID: 7304971
  13. A new and improved method for 3'-end labelling DNA using [alpha-32P]ddATP.
    Gene. 1984 Mar;27(3):309-13 PMID: 6735176
  14. DNA adenine methylation of GATC sequences appeared recently in the Escherichia coli lineage.
    J Bacteriol. 1984 Nov;160(2):586-90 PMID: 6094478
  15. Repair of DNA base-pair mismatches in extracts of Escherichia coli.
    Cold Spring Harb Symp Quant Biol. 1984;49:589-96 PMID: 6397313
  16. Involvement of Escherichia coli mismatch repair in DNA replication and recombination.
    Cold Spring Harb Symp Quant Biol. 1984;49:611-5 PMID: 6397316
  17. Immunochemical detection of minor bases in nucleic acids.
    Science. 1971 Oct 1;174(4004):70-2 PMID: 5120872
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/13.21.7639
Published
1985-11-11
Pages
7639-45
Language
English
Region
England
NLM ID
0411011
PMCID
PMC322076
Subset
IM
Grants
NIGMS NIH HHS · GM012053 · United States
External Links
PubMed source DOI: 10.1093/nar/13.21.7639 PMC Full Text View journal details
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