title: GSM3382514: Cre_Female_3mo_8619_S92: SostERt2 Cre+; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted from grastrocnemius muscle from 1-month-old Sost-CreERT2 and wild type Cre- male mice and 3-month-old Sost-CreERT2 and wild type Cre- female mice  for RNAseq analysis using a fibrous tissue RNeasy kit (Qiagen) and purified using a MinElute RNA cleanup kit (Qiagen) per the manufacturer's instructions.  Muscles from both legs were combined for RNA extraction. Extracted RNA samples were stored at -80°C until use. The concentration and quality of total RNA samples was first assessed using Agilent 2100 Bioanalyzer. A RIN (RNA Integrity Number) of five or higher was required to pass the quality control. Then five hundred nanogram of RNA per sample were used prepare dual-indexed strand-specific cDNA library using TruSeq Stranded mRNA Library Prep Kit (Illumina). The resulting libraries were assessed for its quantity and size distribution using Qubit and Agilent 2100 Bioanalyzer. Two hundred pico molar pooled libraries were utilized per flowcell for clustering amplification on cBot using HiSeq 3000/4000 PE Cluster Kit and sequenced with 2×75bp paired-end configuration on HiSeq4000 (Illumina) using HiSeq 3000/4000 PE SBS Kit. A Phred quality score (Q score) was used to measure the quality of sequencing. More than 90% of the sequencing reads reached Q30 (99.9% base call accuracy).
identifiers.namespace: GEO
identifiers.id: GSM3382514