Subtraction Hybridization in Microplates:An Improved Method to Generate Strain-specific PCR Primers

Summary An improved subtraction hybridization technique was developed and evaluated. The hybridization is performed in a microplate with the subtractor-DNA immobilized in the plate while the probe-DNA is in solution. After hybridization the probe-specific DNA can easily be removed from the microwell and submitted to further analysis. This new technique has been successfully applied to generate several strain-specific PCR-primers for Lactococcus lactis subsp. lactis , Pediococcus spec., Saccharomyces spec. and Listeria monocytogenes .