Rapid and Quantitative Detection of Leifsonia xyli subsp. xyli in Sugarcane Stalk Juice Using a Real-Time Fluorescent (TaqMan) PCR Assay

Figure 3

Sensitivity tests based on 4% ((a) and (c)) or 1.5% ((b) and (d)) agarose gel electrophoresis of the amplicons from plasmid DNA (pMD19T-Pat1) and genome DNA of Leifsonia xyli subsp. xyli (Lxx) using two set primers of qPCR (Pat1-QF/Pat1-QR) ((a) and (c)) and conventional PCR (Pat1-F2/Pat1-R2) ((b) and (d)). The same set of diluted solutions (10⁸–10 copies/μL) of plasmid DNA (pMD19T-Pat1) were carried out in (a) and (b), and the same set of templates (100 ng/μL–10 fg/μL) of genome DNA of Lxx were used in (c) and (d). M1: 20 bp DNA ladder; M2: DL 2000 DNA marker; M3: 100 bp DNA ladder; NC: total DNA (100 ng/μL) extracted from Lxx-free sugarcane stalk juice.

(a)

(b)

(c)

(d)