Typing Method for the QUB11a Locus of Mycobacterium tuberculosis: IS6110 Insertions and Tandem Repeat Analysis

Figure 1

Electrophoresis of a BsmBI-digested large QUB11a fragment amplified from a single Mycobacterium tuberculosis isolate using primers labeled with fluorescent dyes FAM and NED. (a–c) Capillary electrophoresis. (d) Agarose gel electrophoresis. (a) The untreated QUB11a region fragment is outside the limits of the size marker (>1,200 bp) of GeneMapper software (Life Technologies, Carlsbad, CA, USA). Therefore, size calculations generated by the software are unreliable. (b and c) Fragments generated following digestion of QUB11a amplicons with BsmBI. Two fragments, labeled with fluorescent dyes FAM (b) and NED (c), were detected. (d) Agarose gel (2%) electrophoresis analysis of QUB11a loci from four isolates. Lanes 1a and 2a: untreated QUB11a amplicons (>1,200 bp); lanes 1b and 2b: QUB11a amplicons treated with BsmBI; lanes 3a and 4a: untreated QUB11a amplicons (<1,200 bp); lanes 3b and 4b: QUB11a amplicons treated with BsmBI. Lane M, mixed 100 bp and 500 bp DNA ladders.

(a)

(b)

(c)

(d)