GSM2599119: Gra-d3-rep2; Homo sapiens; OTHER
Cells were pelleted, washed twice with ice-cold PBS and kept on ice. Lysis buffer (10 mM Tris-HCl, pH 7.4, 0.5% NP-40, 10% glycerol, 3 mM CaCl2, 2mM MgCl2, 1mM DTT, 1X protease inhibitor cocktail and SUPERase-In) was added and cells were pipetted up and down through 1 ml tips to release nuclei. The nuclei were washed twice with lysis buffer and resuspended in 100ul freezing buffer (50 mM Tris-HCl, pH 8.3, 40% glycerol, 5 mM MgCl2, 0.1mM EDTA, and 4 units of SUPERase-In per mL). 8 x 106 nuclei were used for each library. Nuclear run-on performed as previously described (Franco et al., 2015) library construction were performed as previously described  (Franco et al., 2015) GRO-seq
GEO Accession
GSM2599119