title: GSM6784041: DCP_25_25_3 [ATGGTCCG-CCATAGGG-2]; Homo sapiens; RNA-Seq
library_name: GSM6784041
library_construction_protocol: To conduct high throughput RNA-sequencing, we developed an adapted version of the plexWell (SeqWell, Beverly, MA, USA) plate-based sequencing method to make the method applicable for high throughput in vitro toxicological analysis. In the plexWell protocol, sequencing reactions are prepared in custom 96 well plates, where each well contains a unique oligo barcode. Plates are also assigned a unique oligo barcode. Cells are lysed and cDNA is directly prepared from the cellular lysate. A tagmentation reaction adds the well specific barcode to the cDNA from each well. The tagmented cDNAs are then pooled prior to library preparation, where plate specific barcodes can also be incorporated for ultrahigh sample multiplexing prior to sequencing. For example, 384 wells of treated samples would be processed across 4 custom 96 well plates, where each of the samples receives a well specific barcode and each of the 4 plates receives a plate specific barcode. This leads to each of the 384 samples receiving a unique combination of well and plate barcodes, which can be bioinformatically deconvoluted following sequencing. Libraries were sequenced at Illumina NovaSeq 6000 l at the Advanced Genomics Core.