RiboPure™ RNA Purification Kit (Invitrogen™)

The Ambion™ RiboPure™ Kit is designed for rapid purification of high-quality RNA from tissue samples or cultured cells. The kit combines TRI Reagent™ with glass fiber filter purification to yield exceptionally pure RNA, free of residual proteins and lipids. The kit includes sufficient reagents for 50 purifications from 1–100 mg of tissue or 0.1–20 x 10⁶ cultured cells each.

• Yields the highest purity RNA attainable
• Ideal for the most stringent downstream applications
• Handles problematic samples with ease
• Fast (less than 30 minutes) and easy procedure

RNA isolated using the RiboPure™ Kit 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. Samples are first homogenized in TRI Reagent™, a monophasic solution containing phenol and guanidine thiocyanate, to rapidly lyse cells and inactivate 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 aqueous phase is removed and passed through a glass fiber filter that binds the RNA. The filter is washed, and the RNA is eluted. The procedure is compatible with tissues that have been stored in Ambion's RNAlater™. Total RNA yield is typically 100–500 μg per 100 mg of tissue, depending on the type of tissue.