title: 50:50 mixture of differentiated THP-1 macrophages and K562 cells treated with 1μg/ml of LPS, profiled with ChronoSeq for 12 samples barcoded with unique Time-Tags
design: 50:50 mixture of differentiated THP-1 macrophages and K562 cells treated with 1μg/ml of LPS, profiled with ChronoSeq for 12 samples barcoded with 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 For K562Cells grown in suspension in DMEM with 10% FBS and 1%PEN-STREP at 5%CO2 and 37C.  THP-1 cells were first grown in suspension same as K562 and then treated with 100nM PMA to induce differenciation for 48 hours. This was followed by a 72 hour rest period before harvesting with 10mM EDTA in DPBS. https://kanishkasthana.github.io/ChronoSeq/protocol_for_preparing_cells_AdditionalFiltration.html 1ug/ml LPS manually added to Cell Sample Reservoir 5 minutes after first sample/time-tag 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