FIG. 1.

Overview of Luminex procedure for determining plasmid content. (1) Plasmid-specific regions are amplified by multiplex PCR, using genomic DNA or B. burgdorferi culture as the template. Unincorporated primers and dNTPs in amplified PCR products are removed by treatment with exonuclease I and alkaline phosphatase. (2) Primers that contain an xTAG sequence are utilized in an asymmetric PCR that incorporates biotin-dCTP. (3) Biotinylated products are hybridized with xTAG microspheres that are coupled to antitag sequences and detected by binding of streptavidin-R-phycoerythrin. MFI, mean fluorescence intensity.