GSM8240935: rock, second mixing experiment; Homo sapiens; Mus musculus; RNA-Seq
Cells were dissociated using Trypsin-EDTA, filtered through a cell strainer and resuspended in BD Rhapsody Sample Buffer Libraries were generated on a BD Rhapsody Express machine. Libraries for unmodified samples (unmod) were generated following manufacturers recommendations for the WTA assay. The protocol for v1 BD Rhapsody was used (i.e. a single RPE reaction) and applied to enhanced beads, either with prior RoCKseq modification (rock, rockroi conditions) or without modification (unmod, unmod_roi). rock, unmod_roi and rockroi conditions were generated following the RoCK and ROI protocol. Briefly, the protocol differs from the standard WTA protocol due to the following points: 1) addtion of N1 primer to the RPE PCR (for unmod_roi, rock and rockroi conditions); 2) addition of ROIseq primers to the pool of randomeres during RPE extension (for unmod_roi and rockroi conditions); 3) separate indexing reaction for the samples in which the N1 primer is added; 4) custom sequencing primer