title: GSM3854985: DKO_TE_RNA seq_1; Mus musculus; RNA-Seq
library_construction_protocol: For blastocyst, to separate ICM and TE from E3.5 embryos(Liu et al., 2016), blastocysts were broken using micromanipulator and washed three times in HCZB. The zona pellucidae of blastocysts were digested with 0.5% pronase E (Sigma) for 10-15 mins and washed three times again in HCZB. The embryos were then incubated in Ca2+-free CZB for 30 mins in 37 ℃, and the ICM and TE were separated into single cells by gently pipetting using a pipette with a diameter of 40–60 μm. Each sample was obtained about 10 cells from one embryo for amplification according the cell shape. For placenta, total RNA was isolated from mouse placentae using Dynabeads™ mRNA DIRECT™ Purification Kit (Thermo Scientific™). For blastocyst, the cDNA library were prepared by SPAPK DNA Sample Pre Kit (Enzymatics) for sequencing using an Illumina HiSeq X Ten sequenator. For placenta, RNA-seq library were prepared by VAHTS mRNA-seq V3 Library Prep Kit for Illumina® (Vazyme), and then applied for deep sequencing on Illumina NovaSeq6000 platform.
attributes.tag: GEO Accession
attributes.value: GSM3854985