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. 2014 Jul 5;15(1):565. doi: 10.1186/1471-2164-15-565

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© Zhang et al.; licensee BioMed Central Ltd. 2014

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Experimental workflow for AmpliSeq-RNA based transcript quantification. An aliquot of the custom primer pool, the cDNA sample and a PCR master-cocktail are combined in individual tubes or multititer plate wells. Following pre-amplification the amplicons are tagged with a sample specific molecular barcode followed by pooling and emulsion PCR (ePCR) amplification on nanosphere-beads. After semiconductor sequencing (Ion-Torrent-Proton) reads are mapped to the target sequence library. Read frequencies proportional to transcript abundance are provided in a standard spread-sheet for further analysis (for experimental details see Methods Section).