GSM8030867
Briefly, digested suspensions of 1 × 105 - 1 × 106 cells from tissues were cross-linked with 2% formaldehyde for 10 min and the reaction was terminated by 0.2 M glycine. The pellet was permeabilized and digested with DpnII overnight followed by proximity ligation. Then, DNA under 1000 bp was extracted and sonicated. To enrich ligation events associated with a specific viewpoint, an appropriate amount of sonicated DNA was taken as a template to linearly amplify for 100 cycles using a 5' biotin-labeled probe of the viewpoint of interest. The amplification products were incubated at 95°C for 5 min, immediately cooled on ice to obtain amplified ssDNA and then enriched with Dynabeads M-280 streptavidin. The bead-bound ssDNA was then ligated with adapters QHR-4C libraries were constructed with specific primer pairs (forward primers containing Illumina P5 with sequences near a specific viewpoint and reverse primers containing Illumina P7 with an index and sequences matching the adapter) and then sequenced on the Illumina NovaSeq platform (Annoroad Gene Technology Co., Ltd.).