title: GSM8809368: DR_MG_2022 GEX library - Mammary Gland- Nulliparous; Pregnant; Lactation; Involution;  Regression; Balb; GF; Mus musculus; OTHER
library_construction_protocol: Mammary glands were isolated in 2% FBS DMEM, minced, treated with collagenase type 2 and digested at 37 degrees in a shaking water bath. The small intestines and colons were collected in 2% FBS RPMI and treated with 1mM DTT and 20mM EDTA at 37 degrees for 15 mins then minced and dissociated in collagenase type 2 with constant stirring at 37 for 40 minutes. Spleen and thymus were collected in 2% FBS RPMI and mechanically disrupted. All samples were filtered and washed in 2% FBS DMEM. All organs, other than spleen, underwent red blood cell lysis and additional washing to obtain a single cell suspension. Libraries were prepared according to the 10x Genomics 3'v3.1 protocol for GEX and FBC