Description
TripleXtractor is a ready-to-use reagent for the isolation of high-quality total RNA, as well as for the simultaneous extraction of RNA, DNA and protein. TripleXtractor can be used with a wide variety of samples, including blood, buffy coat, cultured cells, animal and plant tissues, bacteria and yeast. During the one-step homogenization/lysis procedure, the strong lysis capability of this monophasic solution of phenol and guanidine isothiocyanate results in the disruption of cells and dissolving of cellular components, while maintaining RNA integrity. The purified total RNA is free of DNA and proteins and can be used for all common downstream applications such as cDNA Library construction, Nothern Blotting, RT-PCR, in vitro translation, nuclease protection assays, etc.
Principle
During the one-step homogenization/lysis procedure, the strong lysis capability of TripleXtractor results in the disruption of cells and dissolving of cellular components, while maintaining RNA integrity. After addition of chloroform to the extract and subsequent centrifugation, the mixture is separated into an aqueous (upper) phase containing RNA and an organic (lower) phase. The aqueous phase is then transferred to a new tube and the RNA is recovered by Isopropanol precipitation. DNA and/or proteins can be recovered from the sequential precipitation. DNA can be isolated by ethanol precipitation from the interphase, and additional precipitation with isopropanol yields proteins from the organic phase.