For the isolation of extremely high-quality RNA from yeast without the use of enzymatic pretreatments that may alter gene expression profiles. Each Ambion® kit includes sufficient reagents for 50 purifications each from 107 to 108 yeast cells.
• Highest yields of total RNA available • No enzymatic pretreatment required • Includes DNA-free™ Reagents
It has traditionally been difficult to isolate high-quality RNA from yeast because of problems disrupting the cells. Many protocols use enzymatic pretreatments that may affect gene expression patterns. The RiboPure™-Yeast RNA Isolation Kit uses no enzymatic pretreatments and thus does not alter expression profiles. It combines a simple and efficient glass bead/one step phenol–mediated disruption using Ambion's Vortex Bead Beater Adapter (see Accessory Products) followed by a glass fiber filter– based RNA purification step. After DNase treatment with the included DNA-free™ Reagents, the resulting RNA is exceptionally pure containing little to no genomic DNA (less than 0.05%). The kit has been used extensively at Ambion to isolate high-quality RNA from Saccharomyces cerevisiae, Schizosaccharomyces pombe, Pichia pastoris, and Candida albicans (see Figure). The kit provides the highest yield of any method tested from each of these species (see Figure).
Accessory Products: The Kit requires the use of the Vortex Adapter (SKU #AM10024), which turns a standard laboratory vortex into a bead beater. This adapter fits the Vortex Genie® 2 and holds 12 tubes.
Store 1.5 mL Screw Cap Tubes, Filter Cartridges and Collection Tubes at room temperature. Store Lysis Buffer, 10% SDS, Phenol:Chloroform:IAA, Binding Buffer, Wash Solution 1, Wash Solution 2⁄3 Concentrate and Elution Buffer at 4°C. Store Zirconia Beads, 10X DNase I Buffer, DNase 1 and DNase Inactivation Reagent at -20°C.
Yeast Total RNA Isolated Using the RiboPure-Yeast Kit
RNA Yields From 3 Yeast Species Using Ambion's RiboPure-Yeast or Competitors' Kits