GSM4333397: WT_DMSO_R2_HL-60_TempO-Seq; Homo sapiens; RNA-Seq
Cells were washed in PBS and then lysed in BioSpyder BBN buffer (50 µL per well, 15 min, rt). Lysates were flash-frozen and shipped to BioSpyder technologies. Library was constructed at BioSpyder technologies according to company's standard procedures. In TempO-Seq, each Detector Oligo consists of a sequence complementary to an mRNA target plus a universal (i.e. same for every targeted gene) primer binding site. They anneal in immediate juxtaposition to each other on the targeted RNA template such that they can be ligated together. Ligated detector oligos are PCR-amplified using a primer set (single-plex PCR reaction, with a single primer pair for each sample) that introduces both the adaptors required for sequencing and a sample-specific barcode. The barcode sequences flank the target sequence and are inserted appropriately into the standard Illumina adaptors to permit standard dual-index sequencing of the barcodes and deconvolution of sample-specific reads from the sequencing data using the standard Illumina software. All the PCR-amplified and barcoded samples are pooled into a single library for sequencing. Sequencing reads are demultiplexed using the standard sequencing instrument software for each sample using the barcodes to give a FASTQ file for each.
GEO
GSM4333397
GEO Accession
GSM4333397