title: GSM8151982: mESC_6d_nucleus_2; Mus musculus; OTHER
library_construction_protocol: AMAR-seq employs a stepwise strategy to lyse cell and nuclear membranes. Then, the RNA fraction undergoes transcriptome library construction on the chip based on the smart-seq2 protocol. For nuclear , cytosines on the GpC sites in the open chromatin regions are methylated with GpC methylation transferase, M.CviPI, to mark the chromatin accessibility. Subsequent bisulphite conversion converts all normal cytosines to uracil, leaving endogenously methylated CpG and artificially labelled methylated GpC unchanged. Since DNA methylation information occurs predominantly at the CpG site and the endogenous GpC methylation level is less than 2%, it is reasonable to use CG methylation to indicate DNA methylation level. The treated GC methylation marks open regions to obtain bilayer molecular information on the nuclear fraction of the cell, culminating in individual cell tri-omics data. Subsequently, AMAR-seq employs a stepwise strategy to lyse cell and nuclear membranes. Specifically, glycoproteins expressed on the surface of the cell membrane are used to encapsulate the cell in a layer of ConA beads, where the cell is lysed in a hypotonic environment, with the cytoplasm flowing out while the nucleus remains intact. Neighboring electrodes are then energized to drag away droplets of cytoplasmic RNA and leave the nucleus at the hydrophilic site. Then, the RNA fraction undergoes transcriptome library construction on the chip based on the smart-seq2 protocol, including reverse transcription, cDNA amplification, and Tn5 transposase-based library construction. After nuclear membrane lysis, cytosines on the GpC sites in the open chromatin regions are methylated with GpC methylation transferase, M.CviPI, to mark the chromatin accessibility. Subsequent bisulphite conversion converts all normal cytosines to uracil, leaving endogenously methylated CpG and artificially labelled methylated GpC unchanged. Since DNA methylation information occurs predominantly at the CpG site and the endogenous GpC methylation level is less than 2%, it is reasonable to use CG methylation to indicate DNA methylation level. The treated GC methylation marks open regions to obtain bilayer molecular information on the nuclear fraction of the cell, culminating in individual cell tri-omics data. smart-seq2 and scNOMe-seq