Profiling R loop in mouse and human hematopietic cells with low cell input using S9.6 antibody coupled cut&tag
Present R loop profling oftern requires large amout of material, lacking information about R loop in primary cells. To overcome this setback, we delevopment low input method which integrates in vitro amplication capability of T7 promoter and high binding specificity between nanotag and IgH (mouse) Overall design: R loop was first recognized and bond by S9.6 antibody, nanobody T5 which recognize mouse IgH kappa was later used to bind S9.6 and ennable the cutting of chromosome site in the vicinity of R loop loci. Fragmaentated DNA was in vitro trancribed by T7 promoter and reverse transcribed to get double strand DNA library for NGS sequencing.