ALA induced transcriptome and single cell microscopy of M.tuberculosis

Single cell microscopy of porphyeins in vegetative and dormant Mycobacterium tuberculosis. Confocal fluorescence microscopy, life-time measurements, and microspectrofluorimetry. Fluorescence lifetime measurements were performed on a PicoQuant MicroTime 200 confocal scanning system (Pico-Quant GmbH, Berlin, Germany) based on an Olympus IX-71 inverted fluorescence microscope (Japan). SymphoTime® software was used for data collection. Fluorescence spectra were recorded in the confocal mode of the MicroTime 200 system using a Shamrock 163 spectrograph with a Newton DU-970 camera (Andor, UK).

Transcriptomic analysis of cells of M. tuberculosis in a vegetative state and under transition into dormant state upon administration of exogenous ALA. The quality of the resulting libraries was checked using the Fragment Analyzer. Quantitative analysis was performed by qPCR. After quality control and assessment of DNA quantity, the pool of libraries was sequenced on an Illumina NovaSeq 6000 instrument (length of reads - 150 bp on both sides of the fragments). FASTQ files were generated using bcl2fastq v2.20 Conversion Software (Illumina). The quality data string record format is Phred 33. As a result, 1,107,614,502 reads were received. 

This study was funded by Russian Science Foundation grant 19-15-00324.