title: GSM5554369: MII young oocyte untreated 1; Mus musculus; RNA-Seq
library_construction_protocol: 7 µl of soft lysis buffer containing 500 mM KCl (Sigma), 100mM pH 8.0 Tris-HCl (Sigma), 1.35 mM MgCl2 (Sigma), 4.5 mM DTT (Thermo Fisher Scientific), 0.45% Nonidet P-40 (Sigma), 0.18U SUPERase-In (Thermo Fisher) and 0.36U recombination RNase-inhibitor (Thermo Fisher) were added into each PCR tube containing a single oocyte and were incubated at 4˚C for 30 min. Then, the lysate was vortexed at room temperature for 1 min and centrifuged at 1000g for 5 min to allow the nucleus to sink to the bottom of the tube. 6.5 µl of supernatant containing RNA was carefully transferred to a new PCR tube for generating scRNA-seq library. RNA libraries were prepared for sequencing the TruePrep DNA Library Prep Kit V2