title: GSM7866372: Jurkat cells, stimulated, replicate 2, LSV-seq; Homo sapiens; OTHER
library_construction_protocol: Total RNA was purified from cells with the Maxwell RSC 48 instrument using the Maxwell RSC simplyRNA Cells Kit (AS1340, Promega). For LSV-seq library preparation, the targeted primer pool was gradually annealed to input RNA using a touchdown step and first-strand cDNA synthesis was carried out at an elevated temperature of 60°C to maximize the specificity of the RT reaction. Next, second-strand synthesis was performed and the double-stranded DNA then served as a template for linear amplification by in vitro transcription (IVT). The IVT amplified RNA (aRNA) was then fragmented and subjected to an additional RT step to append a second adapter sequence. The resulting cDNA was then amplified by PCR to generate the final sequencing library.