GSM4506239: pCAF; Mus musculus; RNA-Seq
Normal mammary fat pad: Tissue was minced using scissors and placed and disrupted for 25 min at 37oC in a gentleMACS C tube using gentleMACS dissociator, in the presence of enzymatic digestion solution containing 1 mg ml-1 collagenase II, 1 mg ml-1collagenase IV and 70 unit ml-1 DNase. Primary tumor: Tissue was minced using scissors and treated with enzymatic digestion solution containing 3 mg ml-1 collagenase A and 70 unit ml-1 DNase in RPMI 1640 for 20 min at 37oC, with pipetting every 3 min. Lung metastases: Metastases bearing lungs were harvested and placed in gentleMACS C tubes with an enzymatic digestion solution containing collagenase A 1.5 mg ml-1, dispase II 2.5 unit ml-1 and DNase I 70 unit ml-1 in RPMI 1640. The tissue was disrupted for 30 min at 37oC using gentleMACS dissociator. Following dissociation, all tissues were washed, filtered through a 70 μm cell strainer and pelleted by centrifugation at 350 g, 5min, 4°C. 3' end mRNA libraries were prepared for sequencing using a method developed at Dr. Ido Amit's lab, Weizmann Institute of Science
GEO Accession
GSM4506239