QIAwave RNA Mini Kit (250)

From QIAGEN, this kit is designed for purification of total RNA from animal cells, animal tissues, bacteria, and yeast, and for cleanup of RNA from crude preparations and enzymatic reactions such as DNase digestion, proteinase digestion, RNA ligation, and labeling reactions. It can also be used to purify cytoplasmic RNA from animal cells.

The procedure combines the selective binding properties of a silica-based membrane with the speed of microspin technology. Biological samples are lysed and homogenized in a denaturing guanidine-thiocyanate-containing buffer that inactivates RNases to preserve intact RNA. After ethanol addition, the sample is applied to an RNeasy Mini spin column where total RNA binds to the membrane and contaminants are washed away, with RNA eluted in 30–100 µL water. The kit enables preparation of up to 100 µg total RNA per sample and parallel processing of multiple samples in less than 30 minutes. All RNA molecules longer than 200 nucleotides are recovered, providing enrichment for mRNA because most RNAs shorter than 200 nucleotides (such as 5.8S rRNA, 5S rRNA, and tRNAs, which together comprise 15–20% of total RNA) are selectively excluded.

Protocols accommodate up to 1 x 10^7 animal cells using spin technology, up to 1 x 10^6 animal cells using vacuum/spin technology, up to 30 mg of fresh, frozen, or stabilized animal tissue, and up to 5 x 10^7 yeast cells. For bacterial RNA, use in combination with RNAprotect Bacteria Reagent is recommended. Purified RNA is suitable for downstream applications including RNA-seq, RT-PCR and real-time RT-PCR, dPCR, differential display, cDNA synthesis, Northern, dot, and slot blot analyses, primer extension, Poly A+ RNA selection, RNase/S1 nuclease protection, and microarrays. The kit is intended for molecular biology applications, is stored dry at room temperature (15–25°C), and remains stable for at least 12 months after delivery.