A quick, easy and efficient protocol for extracting high-quality RNA from Mycobacterium tuberculosis using a spin column commercial kit.

Publication date: Jul 13, 2023

RNA extraction from Mycobacterium tuberculosis has been a historically challenging task for researchers due to the thick lipids associated with the cell wall of this “notorious” pathogen that is responsible for Tuberculosis (TB) outbreaks. Several studies have successfully extracted RNA from M. tuberculosis using a Trizol reagent combined with organic solvents. Recently, our laboratory has successfully extracted high quality total RNA using a commercial kit from clinical strains belonging to F15/LAM4/KZN, Beijing and F11 strain families and H37Rv laboratory strain by exploiting high speed homogenizer for cell lysis and spin columns for RNA purification. The quality and integrity of the extracted RNA was analyzed and confirmed through the Nanodrop, Bioanalyzer and RNA 3-(N-morpholino) propanesulfonic acid (MOPS) gel electrophoresis. Furthermore, to confirm the integrity of small RNA (sRNA) molecules due to their vulnerability to degradation, the RNA samples were converted to cDNA and sRNAs were amplified and confirmed through PCR. This detailed RNA extraction protocol proposes to carve a new path into TB transcriptome research without the use of organic solvent for downstream purification steps while yielding high quality RNA that can be used to understand M. tuberculosis transcriptome regulation.

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Concepts Keywords
Bioanalyzer Commercial kit
Efficient M. tuberculosis
F15 Protocol
Nanodrop RNA extraction


Type Source Name
disease MESH Tuberculosis
pathway KEGG Tuberculosis
pathway REACTOME Reproduction
drug DRUGBANK Spinosad
disease MESH Infection
disease MESH restriction fragment length polymorphism
drug DRUGBANK Glycerin
drug DRUGBANK Polysorbate 80
drug DRUGBANK Oleic Acid
drug DRUGBANK Dextrose unspecified form
pathway KEGG Cholesterol metabolism
drug DRUGBANK Cholesterol
drug DRUGBANK L-Asparagine
drug DRUGBANK Magnesium sulfate
drug DRUGBANK Tyloxapol
drug DRUGBANK Ethanol
drug DRUGBANK Probucol
drug DRUGBANK Isoxaflutole
drug DRUGBANK Zoledronic acid
drug DRUGBANK Water
drug DRUGBANK Sodium acetate
drug DRUGBANK Edetic Acid
drug DRUGBANK Coenzyme M
drug DRUGBANK Ribostamycin
drug DRUGBANK Ethionamide
disease MESH repression
drug DRUGBANK Iron

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