GSM7061953: biol FAD/Tet1(+/-)_3; Mus musculus; OTHER
Cells were lysed with 600ul of lysis buffer contaning 100mM Tris-HCl, Ph8.5, 5mM EDTA, 0.2% SDS, 200mM NaCl, and 25ul of protease K.Following lysis, RNase A was added and phenol:chloroform:isoamyl alcohol (25:24:1) was used to extract DNA. The Covaris ME220 was used to sonicate gDNA into ~200-300bp fragments. 5hmC was labeled with UDP-6-N3-Glu βGT and biotin. Invitrogen Dynabeads® MyOne™ Streptavidin C1 were used to pull down the biotin labeled 5hmC. 5hmC capture libraries were prepared using the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® manufacturers protocol