title: MietttinenSample7
organism: Mus musculus
description: Protocols: Female mice were grown to 14 weeks of age on normal laboratory conditions before analysis Partial (70%) hepatectomy was performed for two mice of each genotype and liver was allowed to regenerate for 96 hours. Untreated mice (two of each genotype) were used as controls. Frozen liver samples (100-200 mg) were transferred to QIAzol Lysis Reagent (Qiagen) and homogenized using Precellys homogenizer (Bertin Technologies) for 30 sec. After chloroform addition, total was precipitated from the aqueous phase with 1/10 volume of 3M sodium acetate, pH 5.0 and one volume of isopropanol. 15 ug total was fragmented for 3 min at 70 degreesC in fragmentation buffer (Ambion). Fragmentation was terminated by cooling of the sample on ice and addition of EDTA to 17 mM final concentration. 2.5 ul fragmented was then mixed with 0.5 ul 100 mM MgCl2 and diluted to 30 ul with RNAse-free water.