biostudies-arrayexpressKanishk Asthanamixed sampleChronoSeq-Toolshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15964Automated sampling within the ChronoSeq device showed similarly precise tagging: when human, mouse, and mixed reservoirs were processed in sequence, each cell population was exclusively labeled with its designated barcode. Goal of this experiment was to show accurate Time-Tagging of each sample with the ChronoSeq device.biostudies-arrayexpressNucleic Acid Extraction - Oligo-DT beads were used to capture mRNA from lysed Single-Cells inside droplets.Library Construction - ChronoSeq library preparation protocol is very similar to Drop-seq. Beads from all time-tags are pooled together for Reverse Transcription followed by PCR amplification. https://kanishkasthana.github.io/ChronoSeq/protocol_library_preparation_for_dropseq_chronoseq_beads.htmlGrowth Protocol - K562 and EL4 cells grown in suspension in DMEM with 10% FBS and 1%PEN-STREP at 5%CO2 and 37C. https://kanishkasthana.github.io/ChronoSeq/protocol_for_preparing_cells_AdditionalFiltration.htmlSequencing - Illumina NovaSeq X 10B on one lane in PE150 configuration with Custom Read 1 primer. 10% PhiX spike in. Mix primers with Illumina PrimersSample Collection - K562+EL4 (Time Tag 10), K562(Time Tag 11), EL4 (Time Tag 12). Automated cell sampling using ChronoSeq device. Each cell sample is injected with a unique injection of Time-Tagged ChronoSeq beads. https://kanishkasthana.github.io/ChronoSeq/protocol_and_software_for_running_chronoseq_device.htmlOrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesSequence Alignment - mm10-hg19 Drop-seq mixed reference used for alignment. Data processed using ChronoSeq-Tools pipeline. https://github.com/kanishkasthana/ChronoSeq-ToolsData Transformation - UMIs and gene information were used to generate unique transcripts captured and remove PCR duplicates using Drop-seq tools.MetabolomicsUnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq XRNA-seq of coding RNA from single cellsmixed sampleKanishk AsthanafalseK562+EL4 (Time Tag 10), K562(Time Tag 11), EL4 (Time Tag 12) (QC) 215 cells/ μl 3 Time-Tags. ChronoSeq No Stimulus. Only Checking Time-Tags with a Sampling Interval of 7minutesAutomated sampling within the ChronoSeq device showed similarly precise tagging: when human, mouse, and mixed reservoirs were processed in sequence, each cell population was exclusively labeled with its designated barcode. Goal of this experiment was to show accurate Time-Tagging of each sample with the ChronoSeq device.2025-11-03T00:00:00Z2026-05-26T22:02:54.357Z2025-11-03T16:20:13.313ZE-MTAB-15964ERP183599E-MTAB-15927E-MTAB-15928E-MTAB-15956E-MTAB-15955E-MTAB-15947E-MTAB-15946E-MTAB-15894EFO_0002944EFO_0004170EFO_0003789EFO_0005684EFO_0004917EFO_0005518EFO_0003816EFO_0004184