GSM4435253: 10x, abdomen, sample 1; Drosophila melanogaster; RNA-Seq
40 adult male abdomens were quickly dissected in ice cold relaxing buffer (1xPBS, 10 mM EGTA and 10 mM MgCl2) by cutting off the last 2nd and abdominal segment with micro-scissors and hollowing out the abdomen by removing the intestines and male reproductive organs. The dissected tissue was then roughly chopped and transferred to a Dounce homogenizer. The relaxing buffer was replaced with 1.3 ml of nuclei lysis buffer from the Sigma NUC-201 Nuclei isolation kit. Single nuclei suspension from the homogenate was prepared according to the manufacturer's instruction the in the NUC-201 kit. Briefly, a 1.5 M sucrose cushion was used for generating the gradient for density gradient centrifugation. Density gradient centrifugation was performed in 2 ml tubes using a SW55Ti rotor on a Beckman ultracentrifuge. Libraries were generated according to 10X genomics v2 protocol.
GEO Accession
GSM4435253