Development of a modified subtraction hybridization technique and its application for the design of strain specific PCR systems for lactococci

A modified genomic DNA subtraction hybridization approach for the design of strain specific PCR systems was developed and evaluated. The method is based on the magnetic separation of homologous and heterologous hybrids of amplified reference (subtracter) and target (probe) DNA fragments from probe s...

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Bibliographic Details
Published inFEMS microbiology letters Vol. 166; no. 1; pp. 63 - 70
Main Authors Wassill, Lars, Ludwig, Wolfgang, Schleifer, Karl Heinz
Format Journal Article
LanguageEnglish
Published Elsevier B.V 01.09.1998
Subjects
Lactococcus lactis
Magnetic separation
Strain specific PCR
Subtraction hybridization
Lactococcus lactis
Magnetic separation
Strain specific PCR
Subtraction hybridization
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Summary:A modified genomic DNA subtraction hybridization approach for the design of strain specific PCR systems was developed and evaluated. The method is based on the magnetic separation of homologous and heterologous hybrids of amplified reference (subtracter) and target (probe) DNA fragments from probe specific fragments. The sequence data of the latter fragments were used for diagnostic primer design. Strain diagnostic PCR systems were designed and evaluated for Lactococcus lactis subsp. lactis strains WS1678, NIZO 3F27, WS1684 and NIZO L5K5I. At least one of the primers may be used as a strain specific hybridization probe.
ISSN:0378-1097
1574-6968
DOI:10.1016/S0378-1097(98)00312-7