Illumina HiSeq 2000 sequencing; Expression data of CD11b+ cells isolated from WT and C3aR knockout mice after CTX injury
Expression data of CD11b+ cells isolated from WT and C3aR knockout mice after CTX injury
30 ul 10uM Cardiotoxin (CTX) was injected to TA muscle. 24 hours after injury, TA muscle was digested into single cells, and CD11b+ cells was sorted by FACS sortor. RNA was extracted with Trizol reagent, followed by clean-up and DNase I treatment with QIAGEN RNeasy mini kit in accordance with the prescribed protocol provided with the kit. Quality control was performed with Agilent Bioanalyser. the mRNA is enriched by using the oligo(dT) magnetic beads (for eukaryotes). Mixed with the fragmentation buffer, the mRNA is fragmented into short fragments (about 200 bp). Then the first strand of cDNA is synthesized by using random hexamer-primer. Buffer, dNTPs, RNase H and DNA polymerase I are added to synthesize the second strand. The double strand cDNA is purified with magnetic beads. End reparation and 3'-end single nucleotide A (adenine) addition is then performed
Experimental Factor: genotype
C3ar knockout