title: GSM8771702: WT_St10.5_snRNAseq_SubLib1; Xenopus tropicalis; RNA-Seq
library_construction_protocol: Nuclei were isolated from stage 10.5 embryos using previously published discontinuous sucrose gradient protocols followed by a separate centrifugation through 80% glycerol. 200 embryos were homogenized in a 250mM sucrose solution containing glycerol and snap frozen in liquid nitrogen. Homogenates were later thawed on ice, brought to ~2.2M sucrose and centrifuged through a 2.4M sucrose layer at 130,000g for 2 hours at 4oC using a Beckman SW55Ti rotor, followed by a 3400xg centrifugation for 10 min at 4oC through an 80% glycerol cushion containing 0.2 U/ml RNAse and 5mg/ml protease inhibitors. Nuclei were finally resuspended in "nuclear" PBS with 0.5% BSA and 0.2 U/ml RNase inhibitor. DAPI stained nuclei were counted with hemocytometer, then fixed using Parse Bioscience Evercode Fixation Kit (SB1003). Nuclei in DMSO were slow-frozen to -80C overnight. A bar-coded snRNAseq library was prepared using the Parse WT Mini Kit (ECW01010)
identifiers.namespace: GEO
identifiers.id: GSM8771702_r1