GSM2632085: Control Hoxb7 Pulldown 9; Mus musculus; RNA-Seq
Total RNA was purified with Trizol. 4-TU tagged RNA was biotinylated by EZ-link Biotin HPDP (1 mg/ml, Pierce Biotechnology) in N,N-dimethylformamide (DMF, Sigma) for 30min and for 3 hours with EZ-link Biotin HPDP (0.2 mg/ml) in 1 ml of 10 M Tris/HCl, pH7.5, 1mM EDTA for every 100μg of total RNA. RNA was mixed with streptavidin beads (100μl, Miltenyi Biotec Inc), and applied to magnetic columns (Miltenyi Biotec Inc). Biotinylated RNA was eluted with 5% (v/v) β-mercaptoethanol. We finally used RNeasy MinElute Cleanup Kit (Qiagen) to concentrate the biotinylated RNA. Library prepared by using Illumina TruSeq RNA prep kit
GEO Accession
GSM2632085