Effect of LPS on Mus musculus wild type and Tlr4 knockout brown adipocytes
Effect of LPS on Mus musculus wild type and Tlr4 knockout brown adipocytes
Cultured cells were lysed in TRI reagent. RNA was isolated using TRI reagent with additional clean-up using a silica column-based isolation method (ISOLATE II RNA Mini Kit #BIO-52071, Bioline), including DNAse treatment. The RNA integrity of each sample was tested using Agilent 2100 Bioanalyser (RNA Nano (5-500 ng/Î¼L)-Chip service) and samples with RNA Integrity Number (RIN) above 7.5 were deemed suitable for sequencing. 1)mRNA enrichment and purification: rRNA depletion or Oligo dT Selection to enrich the mRNA 2)RNA fragment and reverse transcription 3)End repair, add A and adaptor ligation 4)RCR amplification 5)Single strand separation and cyclization 6)DNA nanoball synthesis 7)Sequencing on BGISEQ platform