Determination of the 3' terminal nucleotide of DNA fragments.

Nucleic acids research ·Vol. 2 ·No. 3 ·1975-03-00 ·Pages 319-25

Olson K, Harvey C

Abstract

A new method for determining the 3'-terminal nucleotide of a DNA strand is presented. Use is made of the fact that one (and only one) 2',3'-dideoxyribonucleotide can be added to the 3'-end of a DNA fragment with calf thymus terminal transferase. Addition of more than one nucleotide analog per strand is impossible due to the absence of a 3'-terminal hydroxyl group. If the terminatind dideoxyribonucleotide contains an (alpha 32p) label, the resulting 3'-blocked strand can be digested by "nearest neighbor" techniques and the original 3'-endgroup determined. Picomole quantities of DNA strands can be labeled and the 3'-end determined.

MeSH Terms

Animals Base Sequence Biochemical Phenomena Biochemistry Cattle DNA DNA Nucleotidyltransferases Deoxyribonucleases Deoxyribonucleotides/analysis Methods Micrococcus/enzymology Nucleotidyltransferases Oligonucleotides Phosphoric Diester Hydrolases Polynucleotides Spleen/enzymology Thymus Gland/enzymology

Chemicals

Deoxyribonucleotides Oligonucleotides Polynucleotides DNA DNA Nucleotidyltransferases Nucleotidyltransferases Deoxyribonucleases Phosphoric Diester Hydrolases

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Olson K

Harvey C

References (12)

12 references, click to expand

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/2.3.319

Published

1975-03-00

Pages

319-25

Language

English

Region

England

NLM ID

0411011

PMCID

PMC342838

Subset

External Links

PubMed source DOI: 10.1093/nar/2.3.319 PMC Full Text View journal details

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Determination of the 3' terminal nucleotide of DNA fragments.

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