Journal: PLoS ONE
Article Title: Design of a new multiplex PCR assay for rice pathogenic bacteria detection and its application to infer disease incidence and detect co-infection in rice fields in Burkina Faso
Figure Lengend Snippet: Sensitivity of the newly described multiplex PCR method compared to the corresponding simplex PCR for each of the targeted bacterial taxon. Every reaction was performed with six samples of corresponding control bacteria at different concentrations. Lane 1: 1 ng/μl, lane 2: 0.5 ng/μl, lane 3: 0.1 ng/μl, lane 4: 0.05 ng/μl and lane 5: 0.01ng/μl, lane 6: water control. a-b: P . fuscovaginae strain UBP735, c-d: B . glumae strain NCPPB 3923, e-f: Sphingomonas strain V1-2, g-h: X . oryzae pv. oryzae strain BAI10, i-j: Pantoea strain ARC10. a, c, e, g, i: simplex PCR with only one primer pair in each case. b, d, f, h, j: multiplex PCR including the five primer pairs.
Article Snippet: Following the optimization, the final conditions for the multiplex PCR protocol were: 2 μl of DNA added to 23 μl master mix comprising 5 μl Hot Firepol Multiplex Mix ready to load 5X (Solis BioDyne,Tartu, Estonia), 1.25 μl (NH4 )2 SO4 (160 mM), 0.2 μl of each primers specific of P . fuscovaginae at 5 μM, 0.2 μl of each primers specific of B . gluma e and B . gladioli at 100 μM, 0.3 μl of each primers specific of Pantoea spp. at 100 μM and 0.3 μl of each primers specific of X . oryzae at 10 μM and finally 0.1 μl of each primers specific of Sphingomonas spp. at 10 μM.
Techniques: Multiplex Assay, Polymerase Chain Reaction