GSM5328179: Lymph Treg - Burn - CD44low - 5_2; Mus musculus; RNA-Seq
Injury-site draining lymph nodes (axillary, brachial, and inguinal) were harvested and immediately placed in ice-cold culture medium, lymph nodes were mechanically minced through sterile 70- μm cell strainers. Cell preparations were washed twice in culture medium and then strained to remove debris, RNA was purified from 50,000 cells from each sample using the RNeasy Micro Kit (Qiagen, CA) cDNA was synthesized using Takara SmartSeq v4 reagents from 1ng of total RNA. cDNA was fragmented by Covaris AFA to ~150bp and adapters ligated using Swift 2S library prep kit from 2ng of cDNA according to manufacturer's protocol.