An effective method of RNA extraction from bacteria refractory to disruption, including mycobacteria.
A high yield, rapid and simple procedure is described for extracting RNA from mycobacteria and other micro-organisms refractory to disruption. The method yielded 20 microg RNA/109 Mycobacterium bovis BCG, more than 10 times greater than our previous method. Intact full length hsp 70 (dnaK) mRNA was detected by northern blotting and quantitated after heat shock by slot blot hybridisation.
Mangan
J A
JA
Department of Medical Microbiology, St George's Hospital Medical School, Cranmer Terrace, London, SW17 0RE, UK.
Sole
K M
KM
Mitchison
D A
DA
Butcher
P D
PD
D001483:Base Sequence; D003902:Detergents / Q000494:pharmacology; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009163:Mycobacterium bovis / Q000187:drug effects / Q000235:genetics*; D012329:RNA, Bacterial / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000032:analysis; D013296:Streptococcus pneumoniae / Q000235:genetics
D003902:Detergents; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; C063842:dnaK protein, E coli