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Sample GSM4193058 Query DataSets for GSM4193058
Status Public on Nov 27, 2019
Title exp1_Exp92
Sample type SRA
Source name PBMC primary cells from donor
Organism Homo sapiens
Characteristics cell type: PBMC primary cells from donor
molecule type: various proteins
Growth protocol Cells at a density of ~1×10^6/mL were fixed in RPMI medium without serum in 1.6% paraformaldehyde (Electronic Microscopy Sciences) for 10 min at room temperature under gentle agitation. Cells to be analyzed for only protein expression were stored in 1X PBS with 10% DMSO at -80°C until antibody staining.
Extracted molecule total RNA
Extraction protocol QIAquick Gel Extraction Kit (Qiagen, 28704).
PCR with Illumina dual index primers
Library strategy OTHER
Library source transcriptomic
Library selection other
Instrument model Illumina MiSeq
Data processing Library strategy: Single cell barcoding by split-pool synthesis via ligation
FLASH-1.2.11 was used to merge the paired end reads.
seqtk-v1.2 was used to convert merged fastq file to fasta format.
The reads were then parsed to identify cell-id and marker-id based on the white-list info in the associated qdata folder, deduplicated based on cell-id and marker-ids and finally chimeric reads were filtered out.
Expression counts were then used to remove putative debris and doublets cells based on the criteria listed in Singlet_settings.txt in the corresponding qdata folder.
Expression counts matrix was then normalized by scaling the counts by row-means and multiplying by a factor of 10.
A github repository for the software used to process the raw-fastqs is available at the following location:
Supplementary_files_format_and_content: Tab separated text files containing jittered expression counts (both normalized and un-normalized) for different markers for each individual cell
The *tar.gz contains various files containing white-list information for cell-barcode ids, anchor-sequence info, marker-sequence info and settings information for removal of cell-debris and cell-doublets. The files in this folder is used to process the raw fastqs for each of the experiment.
Submission date Nov 26, 2019
Last update date Nov 27, 2019
Contact name Maeve Ohuallachain
Street address 4300 Hacienda Drive
City Pleasanton
State/province CALIFORNIA
ZIP/Postal code 94588
Country USA
Platform ID GPL15520
Series (1)
GSE130784 Ultra-High Throughput Single Cell Analysis of Proteins and RNAs by Split-pool Synthesis
BioSample SAMN13390055
SRA SRX7217821

Supplementary file Size Download File type/resource
GSM4193058_exp1_Exp92.tar.gz 1.7 Kb (ftp)(http) TAR
GSM4193058_exp1_Exp92_12cycles_i702_i502_S1_15_6_30_2.JITTERED_forFCS.txt.gz 5.1 Mb (ftp)(http) TXT
GSM4193058_exp1_Exp92_12cycles_i702_i502_S1_15_6_30_2.UNJITTERED_forFCS_normalized_filtered_Chi2Pval_1.0_jittered0.5.txt.gz 17.0 Mb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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