title: GSM5872526: Dis3_MII_cKO_3; Mus musculus; RNA-Seq
library_construction_protocol: polyA RNAs: Each oocyte was lysed in RLT-plus buffer. The oligo-dT beads were used to isolate the polyA RNA. RiboMinus RNAs and ribosomal RNA: with Ovation Solo kit. CUT&Tag: oocytes were collected according to Bench top CUT&Tag V.3 protocol For polyA RNA seq: Single oocyte RNA-seq libraries were prepared according to a published single-cell RNA seq pipeline with subtle modifications (Macaulay et al., 2016). Briefly, the oocytes at desired stages were collected individually into 2.5 ml RLT plus and stocked at -80 °C until all samples were collected. In the beginning of RNA purification, ERCC RNA spike-in mix (ThermoFisher 4456740) was diluted 10^5 fold, and 1 ml of the diluted ERCC mix was added to each lysis. Poly(A) RNA were isolated by oligo (dT) beads, reverse transcribed, amplified and purified. The purified cDNAs were analyzed by Bioanalyzer 2100 to confirm successful amplification range and quality. cDNAs passing a quality test were further constructed into sequencing libraries by Nextera DNA Sample Preparation Kit. For RiboMinus Total RNA-seq: single oocyte libraries were prepared according to the Ovation Solo system, but random hexamer primer is used for reverse transcription. For CUT&Tag sequencing: libraries were constructed according to the protocol Bench top CUT&Tag V.3, with fresh GV oocytes without light fixation.