GSM5598277: Hippocampus_TG-DTMB 3; Mus musculus; RNA-Seq
Brain was extracted after perfusion with cold PBS. Hippocampus and cortex was dissected from brain after brain extraction. Dissected tissues were frozen with liquid nitrogen and kept at -80°C until further use. RNA was extracted with Trizol reagent. Extracted RNA was treated with DNase to remove remaining DNA in extracted RNA. Then RNA was extracted again through phenol:chloroform extraction. Extracted RNA was confrimed of its concentration and purity through Nanodrop and agarose gel electrophoresis by confirming clear two bands of 28S and 18S rRNA. Extracted RNA was treated with 1/10 volume of 3M sodium acetate (pH 5.2) and 2~2.5 volumes of cold 100% ethanol and kept at -80°C until library construction. Among the extracted RNA, 1μg of RNA was used for library construction. RNA-Seq libraries were prepared by using NEXTflex Rapid Directional mRNA-Seq Kit Bundles (Bioo Scientific, TX, USA) and sequenced on Illumina HiSeq 2500 platform (San Diego, CA, USA).