title: Knockdown of Zona Pellucida protein 2 (ZP2) in mouse embryos by Trim-away and morpholino combined
design: Knockdown of Zona Pellucida protein 2 (ZP2) in mouse embryos by Trim-away and morpholino combined
library_name: Sample 9_p
library_construction_protocol: Approximately 80 hours after microinjection of Trim-away cocktail into pronuclear-stage oocytes, embryos were collected from the culture medium (KSOM with amino acids) at the blastocyst stage. Nine (9) blastocysts, zona-intact, were allocated to each sample. Using a molecular barcode, the samples were demultiplexed (DRAGEN BCL Convert 4.3.13) to fastq data and quality controlled (FastQC version 0.12.1; https://www.bioinformatics.babraham.ac.uk/projects/fastqc/). Trimmomatic (PMID 24695404; version 0.39) was used for adapter trimming and read filtering.  Zygotes were cultured in KSOM medium containing amino acids in 4-well plates, at 37 °C under 6 % CO2 in air.  Pronuclear-stage oocytes were microinjected with Trim-away + morpholino cocktail (directed against either ZP2 or GFP) as treatment, or with sole Trim21 mRNA as control. RNA was extracted and purified using Quick-RNA MicroPrep (cat. no. R1051, Zymo Research Europe GmbH). The library preparation of the total RNA was performed with the Watchmaker mRNA Library Prep Kit (7K0078-024) according to the manufacturer's instructions. Total RNA integrity and quality of the library were assessed using a TapeStation4200 (Agilent, Santa Clara, CA, USA). 