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  Indian J Med Microbiol
 

Figure 3: Optimization of different influence factors in specific PCR of P. praeruptorum. M: DNA Marker (containing a mix of 6 individual DNA fragments from top to bottom denoting 600, 500, 400, 300, 200, and 100 bp). A Amplification results using P. praeruptorum identification primer pairs QH-CP19s\QH-CP19a: 1, 2, and 3 indicate A. decursiva; 4, 5, and 6 indicate P. praeruptorum; 7 indicates negative control without DNA template. B. Annealing temperatures: numbers from 1 to 21 indicate annealing temperatures of P. praeruptorum with 48, 49, 51, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67 and 68 °C; numbers from 22 to 42 indicate annealing temperatures of A. decursiva with 48, 49, 51, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67 and 68 °C. C. Cycle numbers: numbers from 1 to 4 indicate cycle numbers of P. praeruptorum with 20, 25, 30, and 35 cycles; numbers from 5 to 8 indicate cycle numbers of A. decursiva with 20, 25, 30, and 35 cycles. D. DNA template dosages: numbers from 1 to 7 indicate DNA template dosages of P. praeruptorum with 42, 21, 10.5, 5.25, 2.625, 1.313, and 0.657 ng; numbers from 8 to 14 indicate DNA template dosages of A. decursiva with 42, 21, 10.5, 5.25, 2.625, 1.313, and 0.657 ng. E. Taq enzyme species: a indicates Taq DNA Polymerase, b indicates Taq Plus DNA Polymerase, c indicates Hot Start Taq DNA Polymerase, 1 indicates P. praeruptorum, and 2 indicates A. decursiva. F. PCR instruments: a indicates LightCycler ®96 System (Roche company), b indicates Professional standard Thermocycler (Biometra company), c indicates MJ research PTC-200 Peltier Thermal Cycler (Bio-Rad company), 1 indicates P. praeruptorum, and 2 indicates A. decursiva.

Figure 3: Optimization of different influence factors in specific PCR of <i>P. praeruptorum</i>.
<b>M:</b> DNA Marker (containing a mix of 6 individual DNA fragments from top to bottom denoting 600, 500, 400, 300, 200, and 100 bp). <b>A</b> Amplification results using <i>P. praeruptorum</i> identification primer pairs QH-CP19s\QH-CP19a: 1, 2, and 3 indicate <i>A. decursiva</i>; 4, 5, and 6 indicate <i>P. praeruptorum</i>; 7 indicates negative control without DNA template. <b>B.</b> Annealing temperatures: numbers from 1 to 21 indicate annealing temperatures of <i>P. praeruptorum</i> with 48, 49, 51, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67 and 68 °C; numbers from 22 to 42 indicate annealing temperatures of <i>A. decursiva</i> with 48, 49, 51, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67 and 68 °C. <b>C.</b> Cycle numbers: numbers from 1 to 4 indicate cycle numbers of <i>P. praeruptorum</i> with 20, 25, 30, and 35 cycles; numbers from 5 to 8 indicate cycle numbers of <i>A. decursiva</i> with 20, 25, 30, and 35 cycles. <b>D.</b> DNA template dosages: numbers from 1 to 7 indicate DNA template dosages of <i>P. praeruptorum</i> with 42, 21, 10.5, 5.25, 2.625, 1.313, and 0.657 ng; numbers from 8 to 14 indicate DNA template dosages of <i>A. decursiva</i> with 42, 21, 10.5, 5.25, 2.625, 1.313, and 0.657 ng. <b>E.</b> <i>Taq</i> enzyme species: a indicates <i>Taq</i> DNA Polymerase, b indicates <i>Taq</i> Plus DNA Polymerase, c indicates Hot Start <i>Taq</i> DNA Polymerase, 1 indicates <i>P. praeruptorum</i>, and 2 indicates <i>A. decursiva</i>. <b>F.</b> PCR instruments: a indicates LightCycler ®96 System (Roche company), b indicates Professional standard Thermocycler (Biometra company), c indicates MJ research PTC-200 Peltier Thermal Cycler (Bio-Rad company), 1 indicates <i>P. praeruptorum</i>, and 2 indicates <i>A. decursiva</i>.