Figure 2. Gel analysis of multiplex polymerase chain reaction (PCR) with mixed autoclaved bacterial culture samples using primer pool I (A) and II (B). A) Lane 1, DNA ladder; lane 2, Bacillus anthracis (BA) + B. thuringiensis kurstaki (BTK), 1:1; lane 3, BA + BTK, 1:5; lane 4, BA + BTK, 1:9; lane5, Yersinia pestis (YP) + Francisella tularensis (FT), 1:1; lane6, YP + FT, 1:9; lane 7, negative control (no template); lane 8, BA-negative control (no template); lane 9, BA-positive control; lane 10, BTK-positive control; lane 11, FT-positive control; lane 12, YP-positive control. B) Lane 1, DNA ladder; lane 2, BA-positive control; lane 3, BTK-positive control; lane 4, BA + BTK, 1:1; lane 5, BA + BTK, 1:5; lane 6, BA + BTK, 1:9; lane 7, FT-positive control; lane 8, YP-positive control; lane 9, YP + FT, 1:1; lane 10, YP + FT, 1:9; lane 11, negative control (no template); lane 12, blank well. Primer pool I contained 7 primer pairs, BA2, BTK1, FT1, YP1, CB1, VA1, and BM1. Primer pool II contained BA4, BTK2, FT2, YP2, CB2, VA2, and BM2. Each primer pool has 1 primer pair for each biological warfare agent of interest. A positive control was run in single PCR with the corresponding primer pair. See Appendix for the conditions of multiplex PCR.
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