High-throughput Eukaryotic Digital Gene Expression (DGE)

Input Requirement(s)
Single cells, cell lysates, or RNA prepared according to our guidelines

Batch Size(s): 96 samples (RNA only), 384 samples, 1152 samples or 3072 samples

Description
We provide digital gene expression (DGE), a form of RNA-seq developed at BTL that is low cost, highly quantitative, and suitable for single cell input. Preparing RNA-seq libraries from limited amounts of RNA template (e.g., single cells) across a large population of samples can be technically challenging and cost prohibitive. To enable efficient, cost-effective transcriptome profiling of thousands of single cells at a time, we have developed a 3’ DGE protocol that converts poly(A)+ mRNA to cDNA decorated with multiple levels of molecular barcodes (Soumillion et al., 2014). This method preserves strand information and enables very high levels of sample multiplexing (via plate and well barcodes). Importantly, the process uniquely marks each individual transcript molecule with Unique Molecular Indices (UMIs), which essentially barcode each input transcript. UMIs allow us to overcome the effects of bias from library construction or amplification steps that affect all other approaches. This method allows for the identification and quantification of transcripts and is well suited for characterizing major patterns of gene expression variation across populations in a cost-efficient manner.