title: GSM2310549: Single.BLA-M-1_RNA-seq; Macaca mulatta; RNA-Seq
library_construction_protocol: Dynabeads mRNA DIRECT kit (Invitrogen) was utilized to extract RNAs from oocytes and embryos. Oocytes or embryos were washed in PBS containing 0.03% PVP and transferred into a RNase-free Eppendorf tube (500 μl) containing 200 μl lysis/binding buffer from the Kit (Invitrogen). Complete lysis was achieved by gently flicking the tube for 2 min. The sample lysis were then temporarily stored at -80 ℃ until RNA extraction. RNA was extracted following manufactures instruction. RNAs were subject to low input RNA-seq library construction based on Tang's protocol (Tang et al. 2010). In case of overloading, pooled RNA were aliquoted into several tubes and each tube was independently amplified, after 20 rounds of PCR amplification, all tubes contained the same stage samples were mixed together and sent to another 9 rounds of PCR amplification, PCR products was purified and the quality of library were checked by qPCR analysis of a set of house-keeping genes, Agilent 2100 bioanalyzer and QUBIT (Life Technologies) were also employed for quality control. The libraries were deep sequenced on Illumina HiSeq 2000 platform using 90-bp paired-end strategy.
attributes.tag: GEO Accession
attributes.value: GSM2310549