- Now includes TRI Reagent®
- Yields the highest purity RNA attainable
- Ideal for the most stringent downstream applications
- Handles problematic samples with ease
The RiboPure Kit is designed for rapid purification of high quality RNA from tissue samples or cultured cells. The kit combines the robust lysis/denaturant, TRI Reagent, with glass fiber filter purification to yield exceptionally pure RNA, free of residual proteins and lipids.
The recovered RNA is suitable for use in all common downstream applications, including cDNA synthesis, real-time and end-point RT-PCR, microarray analysis, Northern blots, and RNase protection assays.
The RiboPure protocol is especially suited for processing samples that are difficult to disrupt, or samples that have large amounts of ribonuclease or other contaminants. The TRI Reagent-based homogenization and extraction steps remove most of the proteins, lipids, and other extracellular contaminants. The RiboPure procedure is fast (<30 minutes) and straightforward (see schematic, Figure 1). Samples are homogenized in TRI Reagent, a monophasic solution containing phenol and guanidine thiocyanate, which rapidly lyses cells and inactivates nucleases. Addition of bromochloropropane (BCP) results in separation of the homogenate into aqueous and organic phases. RNA partitions to the aqueous phase, while DNA and protein remain in the interphase and organic phase. The RNA is isolated from the aqueous phase through binding of the RNA to a glass fiber filter. After two on-filter washes to remove contaminants, the RNA is eluted from the filter with a low salt buffer.
The RiboPure RNA Isolation Kit includes sufficient reagents for 50 purifications. Each purification can accommodate 1–100 mg of tissue or 0.1–20 x 106 cultured cells.
The procedure is compatible with tissues that have been stored in Ambion’s RNAlater®. Total RNA yield is typically 100–500 ug per 100 mg of tissue, depending on the type of tissue.
Trademarks, Patents, Licensing
|
