Description
The RNeasy MinElute Cleanup Kit enables cleanup and concentration of RNA from enzymatic reactions or other samples using specialized RNeasy MinElute spin columns based on silica-membrane technology. The kit can also be used to desalt RNA samples. Up to 45 µg RNA can be purified in a volume as low as 10 µl. Purification can be fully automated on the QIAcube Connect.
The RNeasy MinElute Cleanup Kit provides high-quality total RNA, free from impurities or enzymatic inhibitors, with A 260 / A 280 ratios of 1.9–2.1 (see figure "High-quality RNA"). RNA amounts corresponding to less than one cell (as little as 1 pg) can be concentrated (see figure "Concentration of RNA"). A large amount of RNA (up to 45 µg) can be purified and is suitable for use in sensitive assays (see figure "Reliable cleanup of RNA"). Reaction volumes can be kept small in downstream applications, giving increased reaction efficiency. The high purity RNA allows more of the sample to be used in reactions without inhibition (see figure "Efficient removal of real-time RT-PCR inhibitors").
The RNeasy MinElute Cleanup Kit is designed to purify and concentrate RNA from enzymatic reactions (e.g., labeling, in vitro transcription), RNA isolated by alcohol-precipitation and organic-extraction methods, as well as to desalt RNA samples, and concentrate RNA prepared by silica-membrane procedures (e.g., PAXgene Blood RNA preps). RNeasy technology simplifies total RNA isolation by combining guanidine-isothiocyanate lysis with silica-membrane purification.
Guanidine-isothiocyanate–containing lysis buffer and ethanol are added to the sample to create conditions that promote selective binding of RNA to the RNeasy MinElute membrane. The sample is then applied to the RNeasy MinElute spin column. RNA binds to the silica-membrane, contaminants are efficiently washed away, and high-quality RNA is eluted in water. Enzymatic reactions or crude RNA preps are simultaneously cleaned up and concentrated in less than 15 minutes. In comparison, time-consuming concentration and cleanup by alcohol precipitation can result in loss of RNA, especially from small samples. The RNeasy MinElute procedure is faster than vacuum centrifugation, which only concentrates the sample without removing salts and other impurities. The unique design of RNeasy MinElute spin columns enables concentration of purified RNA to as little as 10 µl for downstream applications such as microarray analysis and real-time RT-PCR. Purification can be fully automated on the QIAcube Connect.
