GSM7070349
The tumor tissues of the LC model and the IP + LC model were homogenized using 1 mL TRIzol reagent (Life Technologies) and total RNA was extracted using the RNeasy Mini Kit (Qiagen). 180ng of total RNA was used for the construction of sequencing libraries. Barcoded oligo-dT based primer (5'-GCCGGTAATACGACTCACTATAGGGAGTTCTACAGTCCGACGATCNNNNNNNNNNCCCCCCCCCTTTTTTTTTTTTTTTTTTTTTTTTV -3' ; (10)N=UMI, (9)C=cell barcode) was used for single-stranded cDNA synthesis and Second Strand Synthesis Module (NEB, #E6111) was used for double-stranded cDNA synthesis. In-house MEDS-B Tn5 transposase was used for tagmentation and libraries were amplified by 10 cycles of PCR using Phusion High-Fidelity DNA Polymerase (Thermo Scientific, #M0530) and following primers (5'-AATGATACGGCGACCACCGAGATCTACACindexGTTCAGAGTTCTACAGTCCGA-3',5'-CAAGCAGAAGACGGCATACGAGATindex GTCTCGTGGGCTCGGAGATGT-3').