Colon,lung(RNA sequencing data)
Firstly, obtain a single-cell suspension and perform activity detection on it. It is recommended to have a cell activity>85% and good monodispersity with low impurity content. Then, prepare and operate the microfluidic channel water in oil droplet library. To ensure the effective capture efficiency of individual cells, it is generally required to control the cell concentration as much as possible within the range of 700-1200 cells/u. During the library preparation process, individual Gel Beads and cells are encapsulated by separate droplets of oil in water to form GEMs. Each Gel Bead has a unique Barcode and UMI sequence, as well as Poly dT primer sequences used to initiate reverse transcription reactions; Subsequently, in the GEM reaction system, cells undergo fragmentation and lysis, releasing mRNA and initiating reverse transcription reaction with Poly dT primer sequences under the action of reverse transcriptase, generating Full cDNA. Then amplify the cDNA and construct a library.