Quantitative considerations for suspension array assays.
A novel format of a multiplex microbead assay that is capable of accurately and reproducibly measuring small changes in 10(8) copies of a nucleic acid is reported herein. The dynamic range of the assay was 32-fold greater than conventional multiplex microbead assays. The assay was modeled mathematically which enabled the accuracy of DNA quantification to be formally analysed using a logical theoretical framework. The assay was also evaluated on RNA extracts taken from stimulated macrophages, to demonstrate that changes in gene expression can be accurately monitored in a biological system. We believe the reproducibility and accuracy of the assay presented herein will be useful for high-throughput, multiplex sensitive detection of nucleic acids over a large dynamic range in a research or diagnostic setting.
Connolly
Ashley R
AR
Centre for Biomarker Research and Development, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD, Australia.
Palanisamy
Ramkumar
R
Trau
Matt
M
D000818:Animals; D005434:Flow Cytometry; D051379:Mice; D008863:Microspheres; D008954:Models, Biological; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009696:Nucleic Acids / Q000737:chemistry; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D009696:Nucleic Acids; D009841:Oligonucleotides