Fig. 4

a and b: Nested real-time PCR of the primary PCR amplicon amplified from pre-processing 30 catapulted cells from a glass slide using LCM. a Quantification cycles show early detection of undiluted primary amplicon compared to the positive control. The positive control was diluted at 10-fold to avoid primer saturation. b Agarose (1.5%, 4 °C) gel electrophoresis showing the 176 bp nested real-time PCR amplification. The amplification was performed by using primers BnMS949bf and 1105br. Lane 1 and 7: 100 bp ladder; Lane 2: Heat-shock; Lane 3: Heat-shock followed by ethanol precipitation; Lane 4: QIAamp DNA Micro kit; Lane 5: positive control; Lane 6: negative control