title: YYTN-1
organism: Rattus norvegicus
description: Protocols: Brain tissue samples were ground into cell powder with liquid nitrogen and transferred to Trizol lysis solution to allow complete lysis of tissue cells. The disrupted tissue samples were centrifuged for 5 minutes at 12000 g at 4°C. The supernatant was then added to 300 μL chloroform/isoamyl alcohol (24:1), sonicated to 1200 g for 8 minutes at 4°C, and centrifuged at 12000g for 8 minutes. After 2 hours, the supernatant was stored in the refrigerator at - 20°C in an isopropanol centrifuge tube containing 600 μL. The supernatant was discarded after centrifugation at 17500g at 4°C for 25 minutes. Finally, the pellet was washed with 0.9 ml of 75% ethanol, suspended upside down, and centrifuged at 17500g for 3 min at 4℃. For total RNA, the supernatant was discarded, dried for 3-5 minutes, and then washed with 30-200μL of RNase-free water. PolyA tail mRNAs were enriched with OligodT magnetic beads. An appropriate amount of interrupting reagent was added to the obtained mRNA, which was allowed to break at a high temperature. Single-stranded cDNA was synthesized using the interrupted mRNA as a template. A double-strand synthesis reaction system is then configured to synthesize double-stranded cDNA. Purification and recovery of cDNA were accomplished with kits, adding base \" A \" to the 3 'end of cDNA and ligate linkers, followed by fragment size selection and amplification by PCR.