GSM9644507: Knock out GV oocyte Replicate 2; Mus musculus; RNA-Seq
For RNA-seq of knock out and wildtype oocytes, single oocytes were extrated as described previously Han et al. (2020) following the protocol by Picelli et al. (2014). GV oocytes were individually snap frozen in 4 ul lysis buffer containing dNTP mix, oligo-dT primer and SUPERase-In RNase inhibitor (Invitrogen). RNA-seq library preparation was performed according to the Smart-seq2 protocol with slight modifications. 10 cycles of PCR pre-amplification were carried out. 1 ng of cDNA was used for tagmentation with the Nextera XT DNA Library Prep Kit (Illumina). Final libraries were amplified and pooled for single-end Illumina sequencing.