GSM4227527: Pachon_PBS_R2; Astyanax mexicanus; RNA-Seq
Single cell suspension from freshly dissected headkidney tissue from cave and surface morphotypes of A. mexicanus were produced by forcing tissue through 40 µM cell strainer into L-15 media (Sigma), containing 10 % water and 5 mM HEPES buffer (pH 7.2) and 20 U/mL heparin (L-90). The strainer was washed once with L-90 and cells were washed once by spinning cells at 500 x g at 4 ºC for 5 mins. Supernatant was discarded and cells were resuspended in 1 mL of L-90 media (L-15 containing 10 % water, 5 mM HEPES (7.2 pH), 5 % fetal calf serum, 4 mM L-glutamin, Penicillin-Streptomycin mix with 10000 U/mL each). Mature Erythrocytes were reduced via flowcytometric sorting (Influx System) of hematopoietic tissue based on scatter characteristics. Sorted cells were washed once in L-90 media  and run again on Influx system to ensure removal of mature erythrocyte cluster and measure cell viability (percentage live cells after sorting: surface fish 82.2 % and cavefish 88.9 %). Single Cell 3' Reagent kits v2 User Guide Rev D (10X Genomics)
GEO Accession
GSM4227527