title: GSM2640588: SCNT_KDM4E_8-cell_replicate1; Bos taurus; RNA-Seq
library_construction_protocol: Pools of 8 eight-cell embryos were washed three times in RNase-free PBS solution, collected in a drop of PBS within 1 μL, and transferred into the lysis buffer immediately on ice. Samples were then centrifuged at 10000 rpm for 30 s at 4°C, and stored at -80°C. Samples were thawed for cDNA synthesis using SuperScript® III (Thermo Fisher Scientific). After free primer removal and 3' poly(A) tailing steps, cDNA was amplified by Ex Taq® HS (TaKaRa) and fragmented by Covaris S220 System. Short fragments are purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments are connected with adapters. The suitable fragments are selected for the PCR amplification. During the Quality Control steps, Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System are used in quantification and qualification of the sample library. Sequencing libraries were made with the fragmented DNA using NEBNext Ultra DNA Library Prep Kit (New England Biolabs) for Illumina according to manufacturer's instruction.
attributes.tag: GEO Accession
attributes.value: GSM2640588