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RNAClean - RNA & cDNA in vitro Reaction Purification


Agencourts RNAClean kit provides a simple, flexible and highly reproducible method for purifying nucleic acid products generated in common enzymatic RT and IVT reactions such as cDNA and cRNA. The kit is formatted for microarray gene expression analysis experiments and is easily performed in manual or automated formats. RNAClean eliminates the need for vacuum filtration or centrifugation, and delivers superior nucleic acid yield and purity.


Key Benefits
Minimize required starting material: use as little as 1 g of total RNA for Affymetrix GeneChip gene expression experiments.
Easy and fast protocol: purification performed in less than 20 minutes.
Application flexibility: high quality, efficient recovery of small to large size nucleic acid fragments.
Formatted for Affymetrix GeneChip and Eberwine-based RNA amplification methods.


High Yield and Consistent Recovery
5 g of input total RNA was isolated and processed in the standard Affymetrix GeneChip single round amplification protocol. RNAClean purification was compared to Qiagens MinElute (cDNA) and RNeasy (rRNA) column-based filtration methods. The results show that RNAClean delivers consistently higher yields of the final cRNA product (Figure 1). RNAClean recovery of RNA fragments with sizes ranging from ~155 bp9.6 kb was assessed using gel electrophoresis (Figure 2 and Table 1). RNAClean showed higher recovery rates of both small and large RNAs.


RNA Quality
RNA purified using RNAClean is free of nucleases and other contaminates that can affect downstream data integ rity. RNA was incubated at varying temperatures following RNAClean purification for up to 2 days and analyzed using an Agilent 2100 Bioanalyzer to detect RNA degradation (Figure 3). There were no signs of degradation even after a 2 hour incubation at 37 C.

Recovery from Low Concentrations
The RNAClean method is a versatile technique that provides efficient recovery from a wide range of starting RNA concentrations. A 1.9 kb RNA fragment was produced via in vitro transcription and purified using RNAClean reagent (Figure 4). The concentration of the RNA transcript prior to purification ranged from ~100 to 1 ng/L. Recovery results demonstrate that RNAClean efficiently recovers nearly 100% down to 13 ng/L, and was capable of slightly over 60% recovery at concentrations as low as 2 ng/L.


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