title: GSM2157698: CUMS 2 PFC Illumina Hiseq mRNA; Mus musculus; RNA-Seq
library_construction_protocol: Twenty-four hours after the mice were surely tested to demonstrate CUMS-induced depression-like behaviors, these depression-like mice and controls were anesthetized by using Isoflurane and decapitated by guillotine. Both sides of the medial prefrontal cortex were quickly isolated and dissected on the ice-cold glass slides. These cortical tissues were placed into the frozen vials that contained RNAlater RNA Stabilization Reagent (QIAGEN, Germany) at 4°C and stored at -80°C for subsequent analysis. Total RNAs from the tissue of medial prefrontal cortex in each mouse were isolated with TRIzol Reagent (Life Technologies, Carlsbad, CA, USA) based on the manufacturer instruction. Total RNA samples placed in the dried ice were delivered to Beijing Genomics Institute (BGI), China for sequencing analysis. RNA samples were done with quality-control by BGI staff using Agilent 2100 Bioanalyzer (Agilent Technologies, USA) with RNA 6000 nano Reagents Port 1. The concentration of total RNA, the value of RNA integrity number (RIN) and the ratio of 28S to 18S ribosomal RNA were measured. The samples with total RNA amount larger than 10 μg, the concentration larger than 200 ng/μl, the RIN larger than 8, and the ratio of 28S to 18S larger than 1.0 were selected for the construction of transcriptome and small RNA libraries, respectively. mRNAs were extracted from total RNA by oligo(dT) beads. mRNAs were randomly sheared into 200 bp fragments that were reversely transcribed into complementary DNA (cDNAs) by random oligonucleotides. Upon end repair, these synthesized cDNAs were purified by using QiaQuick PCR extraction kit and ligated by sequencing adaptors. After the amplification with Illumina PCR Primer Cocktail within 15 cycles of PCR reaction, these cDNAs were size-selected and purified by agarose gel electrophoresis. cDNAs with sizes between 200 and 300 bp were selected for library construction.
attributes.tag: GEO Accession
attributes.value: GSM2157698