Illumina Genome Analyzer paired end sequencing; RNA-seq of first-trimester chorionic villus for understanding the maintenance of early gestation as well as pathogenesis in pregnancy-related disorders
RNA-seq of first-trimester chorionic villus for understanding the maintenance of early gestation as well as pathogenesis in pregnancy-related disorders
First-trimester placentae (six to nine week of gestation, confirmed by the embryo size under detection of ultrasound) were collected in the Second Affiliated Hospital of Zhengzhou University with the approval of Life Science Ethics Review Committee of Zhengzhou University. All patients signed have been informed consent for the collection and use of these tissues. After being separated ex-vivo, the placentae were immediately washed in the ice saline and divided into several parts by a scalpel blade on ice, and then were transformed into cryogenic vials which were filled with 1 ml RNA store beforehand. When disposed properly, the samples were put into 4℃ refrigerator for 24h so as to let the RNA store immerse them. After 24h the RNA store was abandoned and the samples were sopped up using sterile absorbing paper, then the samples were separately collected into new cryogenic vials and were store into -80℃ refrigerator for further investigations. Total RNA was isolated using the RNeasy Plus Mini Kit (Qiagen) according to the manufacturer's instructions. QIAshredder was used for homogenization. The amount of total RNA extracted was quantified on a Nanodrop 2000 spectrophotometer and RNA quality and purity was further evaluated using an Aligent 2100 Bioanalyzer (Aligent Technologies, Santa Clara, CA, US). RNA-Seq was performed by CapitalBio Corporation (Beijing, China).
Experimental Factor: individual
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