Proof of principle for iPS2-10X-seq: timecourse experiment during 2D cardiac differentiation

Proof of principle and optimization experiment on 10X-based-iPS2-seq was performed in two experiments: The first explorative experiment (H001AS5) was performed on two batches of hPSC and hPSC-derived cardiomyocytes at day 23 and pooled together. hPSC were not treated with Tetracycline or primed for this experiment. Tetracycline has been administered throughout the differentiation. All batches were filtered and counted by a hemocytometer with trypan blue to assess vitality (all over 95% vital); then, they were pooled in the same tube before loading the 10X chip. hPSC and hPSC-derived cells were loaded on different days on separate 10X chips G as 16000 cells aiming as target cell recovery of 10000. The second exploratory experiment (H001AS7) is a time course from Day 0 to Day 12. Day 0 hPSC were primed and treated with Tetracycline 5 days prior library preparation. Then Day 2, Day 6 and Day 12 were treated with Tetracycline throughout and 5 day prior cardiac differentation. Sequencing has been performed for H001AS5 on a Illumina NextSeq 500 using two High Output Kits v2.5 (150 Cycles), for H001AS8 on a NextSeq 2000 on P3 Illumina kit 100 cycles. Sequencing run: Read1, 28 cycles; Index1, 10 cycles; Index2, 10 cycles, Read2, 90 cycles.