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Fig. 3 | BMC Microbiology

Fig. 3

From: A rapid and sensitive system for recovery of nucleic acids from Mycobacteria sp. on archived glass slides

Fig. 3

(i, ii and iii): Agarose gel electrophoresis (1.5%, 4 °C) showing the amplification process of 30 M. smegmatis (NCIMB 8548) cells catapulted from Ziehl-Neelsen archived slide using LCM. (i) Multiple displacement amplification (MDA) by using REPLI-g® UltraFast Mini kit (Qiagen). Samples were extracted using 3 different methods to determine an appropriate method for a lower number of cells isolated from archived glass slides. Lane 1 and 6: 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. (ii) 600 bp product of primary PCR using post-MDA mixture as a template which was performed by using touchdown PCR. Lane 1 and 7: 100 bp ladders; 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. (iii) 176 bp product of touchdown nested PCR amplified from primary amplicon. Lane 1 and 7: The 100 bp ladders; 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

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BMC Microbiology

ISSN: 1471-2180

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