Analysis of small RNAs with the Agilent 2100 Bioanalyzer

Isolation of pure and intact RNA is essential, but the concentration and integrity of total RNA preparations is affected by the purification process used. Applications for which RNA will ultimately be used may include reverse transcriptase (RT)-PCR, preparations of targets for microarrays, ribonuclease protection assays, preparations of cDNA libraries and northern blotting. Recently, the importance of low-molecular-weight (LMW) RNAs, such as small interfering RNAs (siRNAs) and small nuclear RNAs (snRNAs), has been outlined in different studies1. In particular, the importance of a new class of small RNA, microRNAs (miRNAs), has been highlighted. Each miRNA is thought to regulate multiple genes, and as hundreds of miRNA genes are predicted to be present in higher eukaryotes, the potential regulatory circuitry afforded by them is enormous. Standard protocols for the isolation of total RNA and mRNA are not optimized for recovering small RNA molecules and may lead to major loss of miRNAs and other small RNAs. Therefore, an efficient, accurate method is needed to determine the integrity and the concentration of total and small RNAs from various sources. The Agilent 2100 Bioanalyzer offers such advantages, coupled with a rapid, automated system (Vitale, D. Comparing performance of the Agilent 2100 Bioanalyzer to traditional RNA analysis techniques. Agilent publication 5980-2206E; 2002; Lightfoot, S. Quantitation comparison of total RNA using the Agilent 2100 Bioanalyzer, ribogreen analysis and UV spectrometry. Agilent publication 5988-7650EN; 2003; Kuschel, M. Analysis of total RNA with the Agilent 2100 Bioanalyzer. Agilent publication 5968-7493E; 2001).