Analysis of total RNA using the Agilent 2100 Bioanalyzer and the RNA 6000

The RNA 6000 LabChip® kit can be used with the Agilent Technologies 2100 bioanalyzer to determine the integrity and measure the concentration of total RNA samples obtained from various mouse tissues and from E. coli. For each sample the data analysis software automatically calculated the ratio of ribosomal RNA peaks and reported the total RNA concentration in ng/μl. The resolution of the RNA separations on the chip was found to be as good or better than that achieved using denaturing agarose gels and the quantitation was comparable to UV absorption measurements. integrates multiple experimental steps. With the introduction of the Agilent 2100 bioanalyzer utilizing LabChip technology from Caliper Technologies Corp., sample handling, separation, and analysis have been integrated in a more automated manner. Chip-based analysis offers several benefits over existing technology including reduced sample consumption, minimized manual intervention, increased analysis speed and data precision, and minimized exposure to hazardous materials. The RNA 6000 LabChip kit can be used with the Agilent 2100 bioanalyzer to measure both the quantity and the integrity/purity of RNA samples. The integration of these two measurements provides a simple and quick assay that consumes minimal sample amounts. Materials and methods Agilent 2100 bioanalyzer instrument and software All chip-based separations were performed on the Agilent 2100 bioanalyzer, which was controlled by dedicated software running on a PC. The Agilent 2100 biosizing software includes data collection, presentation and interpretation functions. Data can be displayed as a gel-like image and/or as electropherogram(s). For each sample, the ratio of the two major ribosomal RNA bands is automatically determined and displayed with the RNA quantitation data on the electropherogram. An additional data evaluation tool is available for data comparison. The Agilent 2100 bioanalyzer contains high voltage power supplies, each of which is connected to a platinum electrode, which allows the instrument to perform multiple and precisely controlled separations. For the RNA applications, the instrument uses fluorescence detection, monitoring the fluorescence between 670 nm and 700 nm.