title: GSM8558898: PAA3_Emo2_ctrl; Mus musculus; RNA-Seq
library_construction_protocol: Third pharyngeal arch artery and surrounding mesenchyme was micro dissected from E12.5 embryos and immediately transferred into a 48 well plate prefilled with 25mM HEPES in DMEM. Once all samples were collected, tissues were transferred to 100µl of 50mg/ml Liberase TM, 0.1mg/ml DNAse-1, and 25mM HEPES in DMEM and incubated at 37°C for 15 minutes with occasional mixing (all pipetting steps with tissue were carried out with 200µl sterile filter tips to reduce loss of cells). Digestion was stopped by adding 1ml of 10% fetal calf serum (FCS) in DMEM. The cell suspension was filtered through a 40µm strainer. Samples were centrifuged for 7 minutes at 300g and room temperature (RT). After carefully aspirating the supernatant, 100µl RBC lysis buffer was added to the pellet, mixed gently, and incubated for 1 minute at RT. The reaction was stopped by adding 1.5ml of 2% FCS in sterile PBS. Samples were centrifuged at 300g for 7 minutes at 4°C. Supernatant was carefully aspirated and the cell pellet resuspended in 100µl of FACS buffer by gentle pipetting. Library construction was performed following manufacturer's instructions (BD Rhapsody™ system mRNA WTA library preparation protocol).
identifiers.namespace: GEO
identifiers.id: GSM8558898_r1