Gene Expression Nebulas
A data portal of transcriptomic profiles analyzed by a unified pipeline across multiple species

Gene Expression Nebulas

A data portal of transcriptome profiles across multiple species

PRJNA521545: Single-cell RNA sequencing of lymphoblastoid cell lines of European and African ancestries

Source: NCBI / GSE126321
Submission Date: Feb 08 2019
Release Date: Jun 09 2019
Update Date: Mar 09 2020

Summary: Here we present single-cell RNA sequencing (scRNA-seq) data of two LCLs: GM12878 and GM18502, produced from the blood of two female donors with European and African ancestry, respectively.

Overall Design: Cells of three samples (the two LCLs and a 1:1 mixture of the two) were prepared separately using a 10X Genomics Chromium Controller and deeply sequenced.

GEN Datasets:
GEND000137
Strategy:
Species:
Healthy Condition:
Cell Type:
Cell Line:
Protocol
Growth Protocol: GM12878 and GM18502 were purchased from the Coriell Institute for Medical Research. Cells were cultured in the Roswell Park Memorial Institute (RPMI) Medium 1640 supplemented with 2mM L-glutamine and 20% of non-inactivated fetal bovine serum in T25 tissue culture flasks. Flasks with 20 mL medium were incubated on upright position at 37℃ under 5% carbon dioxide. Cell cultures were split every three days for maintenance.
Treatment Protocol: -
Extract Protocol: Single-cell sample preparation was conducted according to Sample Preparation Demonstrated Protocol provided by 10X Genomics as follows: 1 mL of cell suspensions from each cell line (day 4, stable phase) was pelleted in Eppendorf tubes by centrifugation (400g, 5min). The supernatant was discarded, and the cells pellet was then resuspended in 1X PBS with 0.04% BSA, followed by two washing procedures by centrifugation (150g, 3min). After the second wash, cells were resuspended in ~500 uL 1X PBS with 0.04% BSA followed by gently pipetting mix 10-15 times.
Library Construction Protocol: Libraries were prepared using the Chromium Controller (10X Genomics, CA) in conjunction with the single-cell 3'v2 kit. Briefly, the cell suspensions were diluted in nuclease-free water according to manufacturer instructions to achieve a targeted cell count of 5,000 for each cell line. The cDNA synthesis, barcoding and library preparation were then carried out according to the manufacturer's instructions. Libraries were sequenced in the North Texas Genome Center facilities on a Novaseq 6000 sequencer (Illumina, San Diego).
Sequencing
Molecule Type: poly(A)+ RNA
Library Source:
Library Layout: PAIRED
Library Strand: Forward
Platform: ILLUMINA
Instrument Model: Illumina NovaSeq 6000
Strand-Specific: Specific
Samples
Basic Information:
Sample Characteristic:
Biological Condition:
Experimental Variables:
Protocol:
Sequencing:
Assessing Quality:
Analysis:
Data Resource GEN Sample ID GEN Dataset ID Project ID BioProject ID Sample ID Sample Name BioSample ID Sample Accession Experiment Accession Release Date Submission Date Update Date Species Race Ethnicity Age Age Unit Gender Source Name Tissue Cell Type Cell Subtype Cell Line Disease Disease State Development Stage Mutation Phenotype Case Detail Control Detail Growth Protocol Treatment Protocol Extract Protocol Library Construction Protocol Molecule Type Library Layout Strand-Specific Library Strand Spike-In Strategy Platform Instrument Model Cell Number Reads Number Gbases AvgSpotLen1 AvgSpotLen2 Uniq Mapping Rate Multiple Mapping Rate Coverage Rate
Publications
Single-Cell Expression Variability Implies Cell Function.
Cells . 2019-12-19 [PMID: 31861624]