Comparison of Agilent 2100 Bioanalyzer and Caliper Life Sciences GX II in Functionality, Total RNA Scoring Algorithms and Reproducibility to Evaluate Total RNA Integrity
Year: 2012
Type: article
Abstract: In the field of molecular biology the ability to determine total RNA integrity is a valuable tool used to assess the suitability of biological samples for downstream analysis using techniques such as microarray profiling, RT-PCR, and genomic sequencing. Using the results of such analyses, RNA degradation status and contamination can be measured and the data used to predict sample performance in genomic assays. Historically, agarose gel electrophoresis was the quality assessment technique used in most molecular biology laboratories but this procedure requires microgram quantities of RNA and the sensitivity of the assay is often dependent on the interpretation of a photographed gel image. With the development of the Agilent 2100 Bioanalyzer, a microfluidics-based platform, samples are now being evaluated more quickly and precisely. This instrument allows researchers to use low amounts of RNA and the Agilent software provides quality assessment metrics such as ribosomal RNA ratios, concentration estimates, a pseudo -gel image, electropherogram traces and an Agilent algorithmic interpretation of the sample RNA integrity (“RIN”-RNA Integrity Number). With a 12 sample per chip limitation and a 30 minute per chip processing time, the Agilent 2100 Bioanalyzer system can be considered relatively low-throughput and may not meet the needs of a laboratory that requires processing of samples on a larger scale. With the development of the Caliper GX II, a high-throughput microfluidics-based platform, 96 samples can be processed in 2.5 hours. In this study, the Agilent 2100 Bioanalyzer and the Caliper GX II have been evaluated to compare total RNA scoring algorithms, reproducibility, and the ability to provide informative RNA quality metrics to help in predicting microarray performance. It was observed that the performance of the two instruments is comparable in functionality and reproducibility but there are notable differences between the RNA scoring algorithms.
Source: Europe PMC (PubMed Central)
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W30685768