title: 50:50 K562:EL4 (QC) at 430 cells/ μl with 10ng/ml TNF-α add 5 minutes after first sample and a sampling interval of 10 minutes. A total of three samples taken with ChronoSeq and tagged with three unique time-tags
design: 50:50 K562:EL4 (QC) at 430 cells/ μl with 10ng/ml TNF-α add 5 minutes after first sample and a sampling interval of 10 minutes. A total of three samples taken with ChronoSeq and tagged with three unique time-tags
library_construction_protocol: 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.html 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.html 10ng/ml TNF-α added to stimulate the cells. Added 5 minutes after first sample. Oligo-DT beads were used to capture mRNA from lysed Single-Cells inside droplets. ChronoSeq library preparation protocol is very similar to Drop-seq. Beads from all time-tags are pooled together, then split into four equal parts for Reverse Transcription followed by PCR amplification. https://kanishkasthana.github.io/ChronoSeq/protocol_library_preparation_for_dropseq_chronoseq_beads.html