Ann Clin Microbiol.  2016 Mar;19(1):20-23. 10.5145/ACM.2016.19.1.20.

An Effective Method of RNA Extraction from Mycobacterium tuberculosis

• Oh TS
• Kang HY
• Nam YS
• Kim YJ
• You EK
• Lee MY
• Cho SY
• Lee HJ

• KMID: 2160081
• DOI: http://doi.org/10.5145/ACM.2016.19.1.20

Abstract

In the RNA-based study, it is important to extract high-quality RNA. However, RNA extraction from Mycobacterium tuberculosis is problematic due to its thick, waxy cell wall rich in mycolic acid, which renders the cells resistant to lysis. Using TRIzol reagent and several powerful bead-beating steps, a high quantity of RNA was obtained.

Keyword

Mycobacterium tuberculosis; RNA extraction; Quantitative reverse transcription polymerase chain reaction

MeSH Terms

Cell Wall
Mycobacterium tuberculosis*
Mycobacterium*
Mycolic Acids
RNA*
Mycolic Acids
RNA

Figure

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  Fig. 1. Ethidium bromide-stained 1% agarose gel of RNA extracted from clinical isolates of M. tuberculosis. All lanes showed 16S and 23S rRNA bands. Lanes 1 to 33: M. tuberculosis clinical isolates; lanes 25, 26 and 27 were identical isolates with lanes 19, 23 and 24 respectively.

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  Fig. 2. Ethidium bromide-stained 1% agarose gel of RNA extracted from M. tuberculosis clinical isolates and strain H37Rv. Lanes 2 to 5: 16S and 23S rRNA bands. Lane 1: M. tuberculosis clinical isolate grown in MGIT; Lane 2: M. tuberculosis H37Rv cultured in Enriched Middlebrook 7H9 Broth; Lanes 3 to 5: M. tuberculosis clinical isolates cultured in Enriched Middlebrook 7H9 Broth; Lanes 6 to 7: M. tuberculosis clinical isolates cultured in MGIT.

Reference

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