organism: Homo sapiens
description: Protocols: The placenta was collected from pregnant women who received cesarean section with explanations and consent regarding participation in research and chorionic villi was collected within 15 minutes after delivery. RNA samples were chemically fragmented into 200-nucleotide-long fragments by 25-second incubation at 94 ℃ in Nature fragmentation buffer (10mM ZnCl2, 10mM Tris-HCl, pH 7). The fragmentation reaction was stopped with 0.5 M EDTA (pH 8.0) and then kept on the ice, and next fragmented RNA for immunoprecipitation was purified by RNeasy MinElute Cleanup Kit (QIAGEN) as instruction. Finally, the size of each sample was checked by Agilent 2100 Bioanalyzer (Agilent). The reverse transcription reaction of input and immunoprecipitated IP RNA samples was carried out with SuperScript Ⅲ (Invitrogen) as manufacturer instruction. Washed in PBS three times to remove maternal blood, the refined villi (about 1 mg) was put into TRIzol® (Life Technologies) and homogenized by a scissor within 15 min after placental delivery. Total RNA was extracted following TRIzol® instruction. Human embryonic kidney cells 293 (HEK293T) cell was incubated in DMEM (Gibco, Invitrogen) containing 4.5 g/L D-glucose and GlutaMAX™ supplemented with 10% FBS and penicillin/streptomycin at 37 ℃ in 5% CO2. Total RNA with RNeasy Mini Kit (QIAGEN) was extracted following the instruction. DNA was removed using an RNase-Free DNase Set (QIAGEN). Enrichment of poly(A) RNA from total RNA of chorionic villi and HEK293T cells was performed two rounds by Dynabeads® Oligo(dT)25 (invitrogen™). The quality of each total RNA and poly(A) RNA was checked by Agilent 2100 Bioanalyzer (Agilent). Strand-specific and 101 bp paired-end RNA-seq libraries were generated with SureSelect Strand Specific RNA (Agilent) following manufacturer's protocol.
attributes.tag: ENA first public
attributes.value: 2018-08-01
attributes.tag: ENA last update
attributes.value: 2018-02-15
attributes.tag: External Id
attributes.value: SAMEA104592341
attributes.tag: INSDC center alias
attributes.value: Maternal-Fetal Biology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan.
attributes.tag: INSDC center name
attributes.value: Maternal-Fetal Biology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan.
attributes.tag: INSDC first public
attributes.value: 2018-08-01T17:01:57Z
attributes.tag: INSDC last update
attributes.value: 2018-02-15T15:30:09Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6507:SFD2_input
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: clinical information
attributes.value: small for date with maternal preeclampsia
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: preeclampsia
attributes.tag: organism part
attributes.value: placenta
attributes.tag: sample name
attributes.value: E-MTAB-6507:SFD2_input
attributes.tag: sampling site
attributes.value: chorionic villus
attributes.tag: sex
attributes.value: female