Bioanalyzer

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. 

Kiewe P et al (2009) Prediction of qualitative outcome of oligonucleotide microarray hybridization by measurement of RNA integrity using the 2100 Bioanalyzer capillary electrophoresis system. Ann Hematol 88 1177-1183. doi 10.1007/s00277-009-0751-5... 

Bioanalyzer can be considered another version of commercial biosensors for off-line analysis. It was developed to have capabilities of complete analysis, short response time, specificity, and sensitivity that allows a quick clinical test. Abbott Vision, Boehringer-Mannheim Reflectron, and Kodak Ektachem DT60 (IBI Biolyzer is the new name) are used for cholesterol measurement in doctors offices. Bioanalyzer consists of biological and transducing component that are not physically connected. The uniqueness of this separation provides the versatility of analysis, i.e., use of disposable and different biological component for multi-components measurements. In authors laboratory, Kodak Ektachem DT60 was used successfully to determine cholesterol in some food matrices as well as in off-line process control. The analysis time was only 10 minutes compared to 1-2 days for the GC and HPLC methods. Complicated... 

Very few immunosensors are commercially available. The commercial immunosensors are either the detector or bioanalyzer types. The PZ 106 immunosensor from Universal Sensors Inc. (New Orleans, LA) has been used as a detector to measure antibody-antigen reaction. Ohmicron (Newtown, PA) developed a series of pesticide immuno-bioanalyzers that have been used in field tests. Pharmacia Biosensor USA (Piscataway, NJ) recently introduced BIAcore immunodetection system. A combination of a unique flow injection device and surface plasmon resonance (SPR) detection technique provides a real time analysis. A carboxylmethyldextran layer added to plasmon generating gold film is a hydrophobic, activatable, and flexible polymer that provides high antibody and low non-specific bindings. System demonstration at the Institute of Food Technologists (IFT) 1994 meeting in Atlanta drew attention of food scientists. It should easily be adapted for food protein characterization. 

Agilent 2100 Bioanalyzer and Protein 200 plus lab-chip kit (Agilent, Palo Alto, CA). 

For the Bioanalyzer, the protein LabChip is prepared according to the manufacturer s instructions. 

The LabChip is then placed in an Agilent 2100 Bioanalyzer and the Protein 200 Plus program is run. 

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.

Emrich, C.A., Tian, H., Medintz, I.L., Mathies, R.A., Microfabricated 384-lane capillary array electrophoresis bioanalyzer for ultrahigh-throughput genetic analysis. Anal. Chem. 2002, 74, 5076-5083. 

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

The so-called micro-total analytical systems (/tTAS) can integrate sample handling, separation, and detection on a single chip [9]. Postcapillary reaction detectors can be incorporated as well [10]. Fluorescence detection is the most common method employed for these chip-based systems. A commercial instrument (Agilent 2100 Bioanalyzer) is available for DNA and RNA separations on disposable chips using a diode laser for LIF detection. In research laboratories, polymerase chain reaction (PCR) has been integrated into a chip that provides size separation and LIF detection [11]. 

Nachamkin I, Panaro NJ, Li M, Ung H, Yuen PK, Kricka LJ, et al. Agilent 2100 bioanalyzer for restriction fragment length polymorphism analysis of the Campylobacter jejuni flagellin gene. J Clin Microbiol 2001 39 754-7. 

LJ, Wilding P. Evaluation of DNA fragment sizing and quantification by tire agilent 2100 bioanalyzer. Clin Chem 2000 46 1851-3. 

The first step consists of RNA extraction and purification. Different criteria can be applied to decide which RNA fraction is to be analyzed, but it is very common to rely on total RNA. Before proceeding to the next step, the yield and quality of the purified RNA have to be determined (44). RNA quantity may be determined, for example, with the Picogreen assay kit (45) or the Agilent Bioanalyzer (46). RNA quality may also be determined using Agilent s Bioanalyzer. If quantity and quality of RNA are within acceptable limits, one can proceed to the next step. 

The application of immobilized sulfite oxidase in the Multipurpose Bioanalyzer (Provesta, USA) has been described by Smith (1987). 1.8 U of the enzyme was entrapped in a gel at the active tip of an oxygen probe. The sensor is suitable for sulfite determination in food samples down to 100 ppm and has a lifetime of 84 days. 

Fig. 4.28. Bench-top Agilent 2100 Bioanalyzer" (Photograph courtesy of Agilent Technologies)...

The advent of microfluidic devices like the Agilent 2100 Bioanalyzer (Agilent Technologies, Sanata Clara, CA) has diverse implications for CE-based protein analysis. This commercially available microchip CE platform is a dedicated instmment that uses manufacturer-supplied reagent... 

The design, manufacture, and operation of the 384-lane tiCAE bioanalyzer likely represents a practical maximum for throughput in microfabricated electrophoresis systems. The fabrication... 

FIGURE 46.10 Design of the 384-lane pCAE bioanalyzer. Channels are grouped into quartets, which share a common cathode reservoir. To save space on the 200-mm diameter substrate, a direct injection scheme is used in place of a cross-injector. The resolution lost by the transition to direct injectors is partially recovered by an increased effective separation length of 8.0 cm. (Adapted from Emrich, C.A. et al.. Anal. Chem.. 74, 5076, 2002. With permission.)... 

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