Rajagopalan M, Boggaram V, Madiraju M V
Department of Microbiology, University of Texas Health Science Center at Tyler 75710, USA.
Lett Appl Microbiol. 1995 Jul;21(1):14-7. doi: 10.1111/j.1472-765x.1995.tb00995.x.
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.
从分枝杆菌中分离总细胞RNA一直是一项劳动强度大的工作，需要大量细胞，用酚 - 氯仿对细胞裂解物进行多次提取，然后进行氯化铯离心。本文报道了一种简单快速的方法，用于从分枝杆菌中分离RNA，所需细胞数量低至1×10⁷个。如此分离得到的RNA在溴化乙锭凝胶上分析时，主要成分包含16S和23S RNA。此外，该RNA用于通过逆转录随后进行PCR扩增hsp65基因的内部片段。
