RNA-seq of mus musculus: KO fully grown oocytes
Collect oocytes from wild-type and Pdp1 knockout mice at 14 days and 30 days post-birth. cDNA were obtained from GV oocytes using the Single Cell Full Length mRNA-Amplification Kit (Vazyme, N712) according to the manufacturers instructions. Briefly, oocytes were collected (10 GV oocytes per sample) in a 0.2-ml PCR tube with a micro-capillary pipette and each sample was added with 3.5 l sample buffer [lysis buffer : RNase inhibitor = 9:1, including 0.2 l of 1:1000 diluted External RNA Controls Consortium (ERCC) spike-in] and incubated with oligo(dT) primer and a deoxynucleoside triphosphate mixture at 72C for 3 min, then Smart-seq2 reverse transcription reactions were performed to obtain cDNA. After the first-strand reaction, the cDNA was pre-amplified using a limited number of cycles (14 cycles). Sequencing libraries were constructed from 5 ng of pre-amplified cDNA using TruePrep DNA Library Prep Kit V2 for Illumina (Vazyme, TD504) according to the manufacturers instructions. Libraries were sequenced on a Illumina NovaSeq X Plus platform with 150 bp paired-end mode.