PoolMC: Smart pooling of mRNA samples in microarray experiments

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Abstract

Background: Typically, pooling of mRNA samples in microarray experiments implies mixing mRNA from several biological-replicate samples before hybridization onto a microarray chip. Here we describe an alternative smart pooling strategy in which different samples, not necessarily biological replicates, are pooled in an information theoretic efficient way. Further, each sample is tested on multiple chips, but always in pools made up of different samples. The end goal is to exploit the compressibility of microarray data to reduce the number of chips used and increase the robustness to noise in measurements.Results: A theoretical framework to perform smart pooling of mRNA samples in microarray experiments was established and the software implementation of the pooling and decoding algorithms was developed in MATLAB. A proof-of-concept smart pooled experiment was performed using validated biological samples on commercially available gene chips. Differential-expression analysis of the smart pooled data was performed and compared against the unpooled control experiment.Conclusions: The theoretical developments and experimental demonstration in this paper provide a useful starting point to investigate smart pooling of mRNA samples in microarray experiments. Although the smart pooled experiment did not compare favorably with the control, the experiment highlighted important conditions for the successful implementation of smart pooling - linearity of measurements, sparsity in data, and large experiment size. © 2010 Kainkaryam et al; licensee BioMed Central Ltd.

Figures

  • Figure 1 Sparsity in a gene's expression profile. Example of a gene showing only one spike (red circle) across 15 samples. The dotted line marks the median value for the samples.
  • Figure 2 Smart pooling process. Schematic showing the pooling process and the utility of a sparse expression profile. The right column shows a gene's expression across 15 samples, with only 1 spike (highlighted dark square). Samples are mixed according to the pooling design in the middle. The columns of the pooling design represent the samples being pooled and the rows represent the microarray chips used to test them. A black square in the pooling design represents the presence of the sample (along that column) on the corresponding chip (along that row). The highlighted column in the pooling design shows the sample that corresponds to the spike. The left column shows the resulting measurements that contain only two significant values (dark squares), those coming from the sample with the spike.
  • Figure 3 Linearity of pooling process. An example of the comparison between the expression data from a synthetic and a multiplex measurement showing data from all 21,505 genes for a single pooled chip
  • Figure 4 Examples of poolMC results. Four examples of decoding performance. (a) Low spike, low noise case, (b) Low spike, high noise case, (c) High spike, low noise case, and (d) High spike, high noise case. For each gene, expression profiles from monoplex result (black square), decoded synthetic result (red open circle), and decoded multiplex result (blue star with lines) are shown. Inset shows the alignment between synthetic and multiplex measurements (green dots) for the gene across the 12 pooled samples. Raw gene expression values are shown.
  • Table 1: Evaluation of poolMC's performance
  • Figure 5 Asymptotic performance of poolMC pooling design. The approximate number of microarray chips needed based on number of samples used for the pooling experiment and the number of spikes expected in the samples.

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CITATION STYLE

APA

Kainkaryam, R. M., Bruex, A., Gilbert, A. C., Schiefelbein, J., & Woolf, P. J. (2010). PoolMC: Smart pooling of mRNA samples in microarray experiments. BMC Bioinformatics, 11. https://doi.org/10.1186/1471-2105-11-299

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