GSM4024715: HFD0-1; Mus musculus; RNA-Seq
Mediobasal hypothalamic were ice-cold homogenized and incubated with HA antibody to specifically select Pomc-RiboTag active translatome. Antibody-tissue homogenates were mixed with Dynabeads for Immunoprecipitation of RNA-HA complexes. Total RNA was purified using a RNeasy Micro Kit (Qiagen) and quantified using a Quant-iT™ RiboGreen™ RNA assay kit (Invitrogen). RNA integrity was assessed on a 2100 Bioanalyzer device (Agilent Technologies) using the RNA 6000 Pico kit (Agilent Technologies). Sequencing libraries were prepared with the NexteraXT kit (Illumina, San Diego, CA, USA) and were sequenced in a HiSeq 2500 (Illumina) obtaining 50 base single read fragments
GEO Accession
GSM4024715