Open AccessA new method for rapid screening of bacterial species‐ or subspecies‐specific DNA probes
Author(s) -
Kook JoongKi,
Kim MiKwang,
Seong JinHyo,
Kim DongKie,
Kim ByungOck,
Park JooCheol,
Kim KackKyun,
Choe SonJin,
Min ByungMoo
Publication year - 2003
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(03)00021-1
Subject(s) - subspecies , genomic dna , fusobacterium nucleatum , recombinant dna , plasmid , microbiology and biotechnology , biology , dot blot , dna , southern blot , hybridization probe , cloning (programming) , bacteria , genetics , gene , porphyromonas gingivalis , paleontology , computer science , programming language
Abstract A simple assay for the rapid screening of bacterial species‐ or subspecies‐specific DNA probes for the random cloning method is presented, involving the use of genomic DNAs as probes and recombinant plasmid DNAs containing genomic DNA digested with Hin dIII as targets. The optimal amount of target DNAs and the concentration of digoxigenin‐labeled genomic DNA probes were 20 ng and 100 ng ml −1 (or 10 ng and 200 ng ml −1 ), respectively. The method was applied to the development of Fusobacterium nucleatum subspecies‐specific probes. Our results showed that four out of 96 probes were F. nucleatum subspecies‐specific, which was confirmed by Southern blot analysis. Our results indicate that the new method can be used for the rapid screening of species‐ or subspecies‐specific probes.