GSM8679120: Patient07,non-MPR; Homo sapiens; RNA-Seq
Biopsy tissue was obtained from patients prior to neoadjuvant chemotherapy combined with PD-1 monoclonal antibody therapy, and preserved in RNAlater (Servicebio, China). The screened specimens would undergo RNA sequencing (RNAseq)Tissues were Total RNA was extracted using the TRIzol reagent (Invitrogen, CA, USA) according to the manufacturer's protocol. RNA purity and quantification were evaluated using the NanoDrop 2000 spectrophotometer (Thermo Scientific, USA). RNA integrity was assessed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). Then the libraries were constructed using VAHTS Universal V6 RNA-seq Library Prep Kit according to the manufacturer's instructions. The transcriptome sequencing and analysis were conducted by Hongsun Co., Ltd (Shanghai, China) ,Biomarkers Technologies Co., Ltd (Beijing, China) and Biotech Co., Ltd. (Shanghai, China). The libraries were sequenced on an llumina Novaseq 6000 and DNBSEQ-T7 platform and 150 bp paired-end reads were generated. Sequencing libraries were prepared, and the quality of the libraries was assessed by Qubit (Thermo Fischer Scientific), 2100 Bioanalyzer (Aligent), and qPCR. Qualifying samples were sequenced by HiSeq (Illumina), and 150-bp paired-end reads were generated at a depth of 20×.