GSM8137708: nmoD30 scRNA-seq; Mus musculus; RNA-Seq
Mouse neuromesodermal organoids were dissociated by Papain dissociation kit (Worthington). In brief, embryoid bodies were incubated in 1 ml Enzyme A along with 20 μl DnaseI at 37℃ for 20-25 min, following by gentle blowing every 5 minutes with wide-bore tips. After washing and filtering, a suitable volume of PBS with 0.04% BSA was added to resuspend the cells to achieve the cell concentration of 2x106 /ml. Single-cell RNA libraries were prepared using DNBelab C4 kit (BGI-MGI) according to the user's guide. The main steps of library preparation include droplet formation, demulsification, reverse transcription, second-strand synthesis, cDNA and oligo product amplification following size selection. Each obtained cDNA pool was divided into two parts. One part of cDNA was prepared by fragmentation, end repair, adaptor ligation and PCR to get a cDNA library for scRNA-seq. Another part of cDNA was used for amplicon sequencing. Corresponding oligo libraries were generated by PCR amplification from oligo products.In order to better capture information on target site, we divided the obtained  cDNA into two portions. One portion was used for normal transcriptome sequencing, while the other portion was used for amplicon sequencing.