title: DRUG-seq data of HepG2 treated with compounds
design: DRUG-seq data of HepG2 treated with compounds
library_construction_protocol: Following treatment, compound-containing media were aspirated and the cells were washed with phosphate-buffered saline (PBS). The cells were then lysed at room temperature for 15 minutes. With the DRUG-seq protocol, there is no requirement for nucleic acid extraction Reverse transcription (RT) primers containing unique molecular identifiers (UMIs) were dispensed into cell lysates using the Echo 665 instrument. An RT mix containing a Template Switching Oligo (TSO) was subsequently added to each well, and the reaction was incubated at 42ì§¸C for 2 hours. After incubation, material from individual wells was pooled and purified using the DNA Clean & Concentrator-100 kit (Zymo Research). The pooled sample was treated with Exonuclease I (ExoI) and further purified, then pre-amplified using DRUG-seq PCR primers. The resulting cDNA was tagmented using loaded Tn5 transposase (Diagenode) and indexed during the final PCR amplification. Final libraries underwent cleaning and size selection with SPRIselect beads. Library quality and concentration were assessed using Qubit (Thermo Fisher Scientific) and Fragment Analyzer (Agilent).