title: GSM7210116: TAMs from MYC-PyMT mouse # 4; Mus musculus; RNA-Seq
library_name: GSM7210116
library_construction_protocol: 7-week-old Control-PyMT and MYC-PyMT mice were lethally irradiated and reconstituted with 3 million total bone marrow cells from WT B6 mice, and treated with low protein diet 2 weeks after. After 6 weeks of dietary switch, mice were sacrificed and tumors were collected for immune cell isolation. TAMs (DAPI-CD45+SiglecF-Ly6G-B220-Ly6C-F4/80+Vcam+) and MTMs (DAPI-CD45+SiglecF-Ly6G-B220-Ly6C-F4/80+Vcam-FOLR2+)14 were sorted based on their surface markers, directly into TRIZOL LS reagents. RNA was extracted using the RNeasy Mini Kit (QIAGEN catalog # 74104) according to instructions provided by the manufacturer. RNA was eluted in 30 µL nuclease-free water. After RiboGreen quantification and quality control by Agilent BioAnalyzer, 500 ng of total RNA with RIN values of 3.3-9.6 underwent polyA selection and TruSeq library preparation according to instructions provided by Illumina (TruSeq Stranded mRNA LT Kit, catalog # RS-122-2102) with 8 cycles of PCR.