A rapid protocol for isolation of RNA from mycobacteria.
Isolation of total cellular RNA from members of mycobacteria has been a labour-intensive task involving large volumes of cells, multiple extractions of cell lysates with phenol-chloroform followed by caesium chloride centrifugation. A simple and rapid procedure is reported for isolation of RNA from mycobacteria using as few as 1 x 10(7) cells. The RNA thus isolated when analysed on ethidium bromide gels contained 16S and 23S RNA as major species. Further, the RNA was used for amplification of an internal segment of hsp65 gene by reverse transcription followed by PCR.
Rajagopalan
M
M
Department of Microbiology, University of Texas Health Science Center at Tyler 75710, USA.
Boggaram
V
V
Madiraju
M V
MV
D001426:Bacterial Proteins / Q000096:biosynthesis; D001483:Base Sequence; D018834:Chaperonin 60; D018833:Chaperonins / Q000096:biosynthesis / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D006146:Guanidines / Q000737:chemistry; D017879:Isothiocyanates / Q000737:chemistry; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics*; D019800:Phenol; D010636:Phenols / Q000737:chemistry; D016133:Polymerase Chain Reaction; D012329:RNA, Bacterial / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000032:analysis; D013010:Sonication
D001426:Bacterial Proteins; D018834:Chaperonin 60; D006146:Guanidines; D017879:Isothiocyanates; D010636:Phenols; D012329:RNA, Bacterial; D012333:RNA, Messenger; C054435:guanidine isothiocyanate; C074119:heat-shock protein 65, Mycobacterium; D019800:Phenol; D018833:Chaperonins