title: GSM4827027: Monkey_16cell_AG_RNA_rep2; Macaca fascicularis; RNA-Seq
library_construction_protocol: SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing (Takara) was used and experiments were performed according to manufacturer's instructions. Briefly, single embryo was incubated in lysis buffer with RNase Inhibitor on dry ice for 1 minutes, and then thawed at room temperature. First-Strand Buffer, SMART-Seq v4 Oligonucleotide, 3' SMART-Seq CDS primer II and RNase Inhibitor was added into the lysed cells and incubated at 72°C for 3 minutes then put on ice immediately. Incubating was performed after adding SMARTScribe II Reverse Transcriptase as following: 42°C for 90 minutes then 70°C for 10 minutes. Without purification, SeqAmp PCR Buffer, PCR Primer II A and SeqAmp DNA Polymerase was added for pre-amplification using thermal cycler for 12 cycles following the manufacturer's instruction. The amplified cDNA was then purified by 1.8× volume of SPRIselect beads and fragmented into an average of 200 bp by sonication using Covaris. Then, the cDNA was prepared into library for sequencing using  NEBNext UltraTM II DNA  Prep Kit for Illumina (NEB, E7103) according to manufacturer's instruction.