title: GSM2170016: Pro-cap Macrophage; Mus musculus; OTHER
library_construction_protocol: ESC and macrophage were incubated in swellinig buffer (10mM Tris-Cl pH7.5, 2mM MgCl2, 3mM CaCl2) , and Cells were lysed in lysis buffer (swelling buffer with 0.5% IGEPAL and 10% glycerol). The resultant nuclei were washed once again with lysis buffer and then re-suspended in 100ml of freezing buffer (50mM Tris-Cl pH 8.3, 40% glycerol, 5mM MgCl2, 0.1mM EDTA). The cDNA products were separated on a 10% polyacrylamide TBE-urea gel and only those fragments migrating between 100-300bp were excised and recovered by gel extraction. Next, the first-strand cDNA was circularized by CircLigase (Epicenter, Cat# CL4115K) and relinearized by APE1 (NEB, Cat# M0282L). Finally, cDNA template was amplified by PCR using the Phusion High-Fidelity enzyme (NEB, Cat# M0530L) according to the manufacturer's instructions. The oligonucleotide primers oNTI200 and oNTI201 were used to generate DNA library for deep sequencing and the primer sequences were described in previous paper.