RNase free water

Highly purified water has become an important reagent in molecular biology. Experiments that involve the use of reverse-transcriptase PCR (RT-PCR), plasmid preparation, construction of cDNA libraries, siRNA, shRNA, miRNA, tRNA and Northern blotting as well as all experiments involving RNA\DNA that require ribonuclease free water. In general, all RNA preparations from cells and tissues are complicated by the presence of RNases. The use of ultrafiltration together with autoclaving allows for the preparation of “RNase-free” water. This process is an effective alternative to DEPC treatment that delivers high purity RNase-free water..

RNases that can be present in water or reagents used during performing molecular biology experiments degrade RNA. Some of them are exonucleases that digest the RNA oligo nucleotides starting from one end of the nucleotide strand and progress along the molecule. Others are endonucleases that begin their digestion process at defined points along the RNA molecules. RNase A has a broad substrate range and degrades single-stranded, double-stranded, and cyclic RNA. Therefore RNase A is often used as a control enzyme to test the quality of “RNase free” water in a RNA stability test.

RNase activity was tested during and after the purification and bottling process by dissolving RNA oligo nucleotides in the water to be tested. A total of three RNA oligonucleotides were dissolved together in RNase free water produced by Biosynthesis and stored at room temperature for several days followed by analysis using mass spectrometry and HPLC to determine the presence of the unmodified oligonucleotides. The results for the analysis are shown below. Notice that RNA oligonucleotides that were stored in ultrapurified TOC 7 18.2 megaohm water for 6 days at room temperature are still intact and remained undigested.

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