Bioanalyzer 2100 From Agilent

The 2100 Bioanalyzer from Agilent is based on the microfluidic Lab-on-a-Chip technology. This technology utilizes a network of channels and wells that are etched onto glass or polymer chips. Different types of chips are available depending upon what you need to analyze: DNA, RNA, proteins. Every chip contains 12 wells/channels, every of them for one sample. The sample volume for the analysis is 1ul. The samples are injected into the separation channel and their components are electrophoretically separated. Pressure or electrokinetic forces move picoliter volumes in a finely controlled manner through the channels. The microchannels are filled with a sieving polymer and fluorescent dye. The dye intercalates into nucleic acids (DNA, RNA) and thus, allow their detection. The components are detected by their fluorescence and translated into gel-like images (bands) and electropherograms (peaks). All the samples are analyzed with high accuracy and reproducibility in only 30 minutes.

In our department, we use the Bioanalyzer 2100 for DNA and RNA evaluation. For DNA, we verify optimal PCR conditions by running PCR products on the chip and evaluating primer-dimer formation, as well as the size and concentration of PCR amplicon. We especially use the Bioanalyzer 2100 for RNA evaluation. We need to evaluate very well the quality of RNA because this is essential for confidence in gene expression analysis using real-time PCR and microarray technology. The Bioanalyzer 2100 is the only apparatus which discriminates good RNA from degraded RNA or RNA contaminated with DNA or proteins. We routinely use a marker for RNA which is 25 nucleotides in length; this marker provides a good means of comparison between runs. The criteria we look for in good quality RNA are a ratio of 1.5 between ribosomal RNAs (28S and 18S), and an homogenous base line: degradation of total RNA is easily detected by a shift in the RNA size distribution towards smaller fragments and a decrease in fluorescence signal of ribosomal peaks.

This instrument requires a PC and a printer. The Agilent 2100 expert software is complex, but very user-friendly. With this software you could automatically quantify individual DNA fragments against internal standards. For RNA analysis, the software integrates the messenger RNA smear and calculates the percentage of ribosomal RNA contamination. Assurance of high RNA quality allows us to work faster and more efficiently by reducing the number of negative control reactions needed for each experiment. Compared to the competition, it’s the best.

The major advantages of the Bioanalyzer 2100 are: Reduced manual labor; fast, accurate and reproducible results; automated data acquisition and analysis; digital data for easy archiving; minimized exposure to hazardous material and reduced waste volume. In conclusion, I could say that this instrument is necessary for all departments of molecular biology that routinely study nucleic acids.