title: Illumina NovaSeq 6000 sequencing; Liquid Biopsy-Based Single-Cell Transcriptome Profiling Characterizes Heterogeneity of Disseminated Tumor Cells from Lung Adenocarcinoma
design: Liquid Biopsy-Based Single-Cell Transcriptome Profiling Characterizes Heterogeneity of Disseminated Tumor Cells from Lung Adenocarcinoma
library_construction_protocol: Malignant pleural effusion sample were obtained from lung cancer patients in Shanghai Chest Hospital and Shanghai Municipal Hospital of Traditional Chinese Medicine with written informed consent. The clinical sample collection and experiment were carried out in accordance with guidelines and protocols that approved by the Ethics and Scientific Committees of both hospitals. MPE was firstly filtered by a membrane to remove cell cluster, followed by centrifuging. Red blood cell lysing buffer (BD) was then added to the cell pellet for lysing red blood cells. The nucleated cells were washed with HBSS, followed by leukocyte depletion. Cell capture and cDNA synthesis using single cell 3 'Library and Gel Bead Kit V3 (10xGenomics) and Chromium Single Cell B Chip Kit (10xGenomics) Single-cell RNA-seq libraries were constructed using Single Cell 3â€™ Library and Gel Bead Kit V3