alias: GSM3681518
title: GSM3681518: CITE-Seq RNA: BM-MNC; Homo sapiens; RNA-Seq
library_construction_protocol: For the bulk experiment:  CD4-/CD8+/CD3+CD69+ and  CD4-/CD8+/CD3+CD69- cells were sorted into wells containing RLT lysis buffer using a SONY SH800 sorter. RNA was isolated by performing a 2X Ampure bead cleanup. RNA was resuspended into RT mix and cDNA synthesis and amplification was performed as described in dx.doi.org/10.17504/protocols.io.nkgdctw. Briefly, we used a modified version of Smart Seq 2 protocol that allows for sequencing of the 3' end of each transcript. For the scRNA/CITE-seq experiment: a panel of 25 antibodies (ADTs) was used to stain the cells according to the CITE-seq protocol. To identify cell doublets, we split cells into 10 aliquots and stained each aliquot with a unique hashing (HTOs) antibody.  Samples were washed and recombined and load on a 10genomics chip. 3prime 10x genomics Version 2 was ran and the manufacturers protocols was followed for cDNA library generation. To recover HTOs and ADTs we followed the CITE-seq protocol found here: https://cite-seq.com/protocol/ CITE-seq (10x genomics 3prime Version 2)
identifiers.namespace: GEO
identifiers.id: GSM3681518