Detection of subpicogram quantities of specific DNA sequences on blot hybridization with biotinytated probes | Zendy

Victor Chan | Zendy; K A Fleming | Zendy; J O McGee | Zendy

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Detection of subpicogram quantities of specific DNA sequences on blot hybridization with biotinytated probes

Author(s) -

Victor Chan,

K A Fleming,

J O McGee

Publication year - 1985

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/13.22.8083

Subject(s) - biology , southern blot , hybridization probe , dna , nucleic acid thermodynamics , molecular probe , microbiology and biotechnology , dot blot , base sequence , genetics , computational biology

A sensitive method for detecting biotinylated DNA probes on dot and Southern blots is described which is based on the principle outlined by Leary et al (1). This system has two main components: detection of biotinylated DNA by a two-step procedure with streptavidin and poly(alkaline phosphatase); and blocking background with Tween 20. 32fg and 80fg of lambda phage DNA was detected on dot and Southern blot hybridizations respectively. 150fg of beta-globin was detected on Southern blots of genomic DNA. This method is fast, reproducible and can detect single copy genes in 0.25 micrograms genomic DNA on Southern blots.

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