Combined sequencing of mRNA and DNA from human embryonic stem cells

• Main Authors: Florian Mertes, Heiner Kuhl, Wasco Wruck, Hans Lehrach, James Adjaye
• Format: Article
• Language: English
• Published: Elsevier 2016-06-01
• Series: Genomics Data
• Subjects: Next generation sequencing; RNA and whole-genome sequencing; Ultra-low input sequencing; Single cell; Embryonic stem cells
• Online Access: http://www.sciencedirect.com/science/article/pii/S2213596016300587

Combined transcriptome and whole genome sequencing of the same ultra-low input sample down to single cells is a rapidly evolving approach for the analysis of rare cells. Besides stem cells, rare cells originating from tissues like tumor or biopsies, circulating tumor cells and cells from early embryonic development are under investigation. Herein we describe a universal method applicable for the analysis of minute amounts of sample material (150 to 200 cells) derived from sub-colony structures from human embryonic stem cells. The protocol comprises the combined isolation and separate amplification of poly(A) mRNA and whole genome DNA followed by next generation sequencing. Here we present a detailed description of the method developed and an overview of the results obtained for RNA and whole genome sequencing of human embryonic stem cells, sequencing data is available in the Gene Expression Omnibus (GEO) database under accession number GSE69471.