title: GSM4774367: PEM22_2 scRNA-seq; Homo sapiens; RNA-Seq
library_construction_protocol: Freshly isolated individual CTCs were selected using a micromanipulator and introduced into a 0.5ml tube with 7ul of 1x lysis buffer (Takara, Cat#634891) containing recombinant RNase inhibitor (0.4U/ul; Takara, Cat#2313A), snap frozen and maintained at -80oC until processing. Single cell lysates were transferred into 96-well plates (Eppendorf, 951020401), and RNA extraction and clean up were carried out using Agencourt RNAClean XP SPRI beads (Beckman Coulter, A63987). Prior to library construction using the Nextera XT library Prep kit (Illumina, FC-131-1096), two steps of quality control were done to evaluate the concentration of each single cell, using Qubit dsDNA high sensitivity assay kit (Life Technologies, Q32854), according to the manufacturer's instructions. Single cell CTC libraries were prepared using a modified Smart-Seq2 protocol (PMID: 30388456). Combined libraries were sequenced on a NextSeq 500 sequencer (Illumina) using the 75 cycles kit, with paired-end 38-base-reads and dual barcoding.