GSM8207341: IL-36Ra-cKO-1; Mus musculus; RNA-Seq
Lyse the tissue using Trizol, centrifuge and transfer the supernatant into an EP tube containing chloroform/isoamyl alcohol (24:1) and mix well. After centrifugation, transfer the supernatant into a new EP tube containing isopropyl alcohol and mix well, then let it stand for 2 hours at (-20°C). After washing, wash 3 times with 75% ethanol. Finally, discard the supernatant and absorb the remaining liquid. After drying, add 20-200 μL DEPC water to dissolve the precipitate. After quality inspection of the RNA, the secondary structure is opened through denaturation, and oligo(dT) magnetic beads are used to enrich the mRNA. Then a fragmentation reagent is added to fragment the mRNA, a reaction system is prepared, and the reaction program is set to synthesize one-strand and two-strand cDNA. Repair the double-stranded cDNA end, add an A base to the 3' end, and connect the adapter to the cDNA. PCR amplification is then performed, and uncirculated linear DNA molecules are digested. The single-stranded circular DNA molecule is then replicated through rolling circles to form a DNA nanoball (DNB) containing multiple copies. The obtained DNBs are added to the mesh holes on the chip using high-density DNA nanochip technology, and sequenced through combined probe anchored polymerization technology (cPAS).