Agilent 2100 Bioanalyzer electrophoresis

Gottwald, E., Muller, O., Polten, A., Semiquantitative reverse transcription-polymerase chain reaction with the Agilent 2100 Bioanalyzer. Electrophoresis 2001, 22(18), 4016 1022. 

We typically use RNA purification by the Trizol method (Invitrogen, following the manufacturer s instructions), which has the advantage over column-based methods that it can purify small amounts of RNA and retain miRs. Purified RNA is dissolved in RJNAse-free water and stored at —80°. RNA quality is assessed on an Agilent Bioanalyzer (Agilent Technologies) or by gel electrophoresis. 

Where the quantity of RNA is limiting, the Agilent bioanalyzer is an extremely useful tool for assessing both the quality and quantity of total RNA. The system is based on capilliary electrophoresis and can be used on samples down to 50 ng mass (for the nano assay) or 25 ng (for the pico assay). 

Fig. 1. Purified proteins imaged using (A) Agilent 2100 Bioanalyzer and (B) traditional sodium dodecyl sulfide-polyacrylamide gel electrophoresis. (A) Note that the lowest two bands and the uppermost band are internal standards for sizing and quantitation. (B) 3-15% polyacrylamide gel stained with GelCode Blue. In both panels, the predicted size of the protein is labeled at the top of each lane.

The building block synthesis is monitored with a microfluidic electrophoresis using the DNA 12000 kit and the Bioanalyzer instrument from Agilent according to the manufacturer s instructions (Fig. 2). This analysis can also be performed with an agarose gel. 

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