Amplification of Mycoplasma iowae using polymerase chain reaction.
Based on sequence data of Mycoplasma iowae recombinant DNA probe, pMI-12, two 25-base primers were synthesized for use in the polymerase chain reaction (PCR). An M. iowae species-specific 299-base pair product was amplified by the primers. An annealing temperature of 58.5 C was critical for detecting all members of this heterogeneous species while maintaining specificity of the M. iowae PCR. The minimum amount of target DNA detected by M. iowae PCR was 1 pg, which was 1000 times more sensitive than the dot-blot assay using M. iowae recombinant DNA probes.
Zhao
S
S
Department of Epidemiology and Preventive Medicine, School of Veterinary Medicine, University of California, Davis 95616.
Yamamoto
R
R
D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial