title: GSM4817931: Healthy heart H8 - Atrium; Danio rerio; RNA-Seq
library_construction_protocol: The heart was dissected from the fish and transferred into cold HBSS. The dissection included the atrium, ventricle and bulbus arteriosus, except for samples in which only the atrium or the ventricle was isolated. A needle and a syringe filled with cold HBSS were used to pierce into the lumen of the heart and thoroughly wash away most of the erythrocytes in the tissue. Afterwards, the tissue was opened carefully with forceps, and the heart tissue was incubated at 37 °C for 30 min in 500µl HBSS containing Liberase enzyme mix (Sigma-Aldrich, 0.26 U/mL final concentration) and Pluronic F-68 (Thermo Fisher Scientific, 0.1%), while shaking at 750 r.p.m. with intermittent pipette mixing. After most of the tissue was dissociated, the reaction was stopped by adding 500µl cold HBSS supplemented with 1% BSA. The suspension was centrifuged at 250g at 4°C and washed two times with 500µl cold HBSS containing 0.05% BSA, then filtered through a cell strainer of 35µm diameter. The quality of the single cell suspension was then confirmed under the microscope, and cells were counted prior to scRNA-seq library preparation. Library were constructed following the manufacturer's protocols (10X Genomics)