title: scRNA-seq of Drosophila pseudoobscura third instal larva testis rep2
design: Each adult sample used 20 pairs of testes from 5- to 7- day-old flies and each larval sample used 20 pairs of testes from third instar (L3) larvae. Testes were dissected in cold PBS and immediately digested in 200 ul lysis buffer (HBSS+5%FBS+2mg/ml collagenase IV) for 10 min at room temperature. After digestion, cells were filtered through 30 m filter and centrifuged 6 min at 600x g. Cells were washed by 200 ul cell suspension buffer (HBSS+5%FBS+0.04%BSA), centrifuged 6 min at 600x g for 2 times and finally resuspended with 100 ul suspension buffer. 5 ul cell suspension were mixed with trypan blue dye to examine cell viability and cell density. Finally, cell concentration is 1000-1200 cells/l.
library_name: larva2