GSM6912706: Hemin-IL-10-2; Mus musculus; RNA-Seq
RNA is extracted from the sample by standard extraction method, and then the RNA sample is subject to strict quality control. The quality control standards mainly include the total amount of RNA, purity (whether there is DNA or protein contamination) and integrity. Qubit RNA HS is used to control the total amount and purity of RNA; The 2100 RNA Nano/Pico kit is used for quality control of RNA integrity. The library construction process is as follows: RNA template chain is reverse transcribed into cDNA. The second strand is synthesized and added to dUTP connector. A is added to the end, sequencing connector is added to both ends. PCR amplification and SPRI purification are performed. After the construction of the library, the preliminary quantification is carried out first, the library is diluted, and then the Agilent 2100 DNA 1000 kit is used to detect the distribution of library fragments. The  concentration of the library is accurately quantified by qRT-PCR to ensure the quality of the library. Finally, the target region library of double chain is denatured, cyclized and digested to obtain single stranded circular DNA. Single stranded circular DNA is amplified by RCA (Rolling Circle Amplification) to obtain DNA nanospheres (DNB, DNA Nano Ball). After the construction of the library, Qubit is used for quantitative quality control. After the library is qualified, computer sequencing is carried out.