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RNA & cDNA Purification Using RNAClean

recovery from a range of starting RNA concentrations. This can be important as the yield of in vitro produced RNA requiring purification can vary widely from application to application. We produced a 1.9kb RNA fragment through in vitro transcription and purified it using RNAClean (Figure 2). The concentration of the RNA transcript prior to purification ranged from over 100ng/l to 2 ng/l. RNAClean shows nearly 100% recovery from reactions containing RNA at concentrations as low as 12ng/l and over 60% recovery from starting RNA concentrations as low as 1ng/l.


Low Volume Elution
RNAClean allows elution in small volumes, which can serve as an effective means to concentrate the cDNA or RNA for downstream application flexibility. We purified a 1.9kb transcript using RNAClean with varied elution volumes from 10 to 40l of water (Figure 3). The percent recovery was greater than 80% for all elution volumes tested.


RNA Quality
RNAClean-purified RNA is free of nucleases and other contaminates which could effect performance in downstream applications. Overall quality of RNA purified with RNAClean from an in vitro transcription reaction was assessed using the Agilent BioAnalyzer 2100. Before testing, the RNA was incubated at either 4C for 2 days, at room temperature for 2 days, or 37C for 2 hours. No detectable degradation was observed in any of the samples tested (Figure 4).

We also tested RNAClean-purified RNA for contamination with impurities that inhibit downstream reactions such as reverse transcription and PCR. RNA transcripts purified with LiCl precipitation or RNAClean were used as templates for first strand cDNA sy
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