title: GSM7413356: IVO MII-1; Macaca fascicularis; RNA-Seq
library_name: GSM7413356
library_construction_protocol: The oocytes were lysed in 72℃ and then put back on ice. Then the samples were spined down and reverse transcribed to cDNA based on polyA tail. Switch the template at 5' end of the RNA and amplify the full-length cDNA by PCR. Quality control of the amplification products were performed using Agilent 2100 bioanalyzer instrument.  The library was constructed following the Tagmentation-based library construction protocol. The double stranded PCR products were purified using the Agencourt AMPure XP-Medium kit and quantified by Agilent 2100 bioanalyzer. Next, the DNA were heat denatured and circularized by the split oligo sequence. The qualified single strand circle DNA were formatted as the final library. Finally, the library was amplified to make DNA nanoballs (DNB) which were loaded into the patterned nanoarray and the reads of paired-end 100 bases were generated in the way of sequenced by combinatorial Probe-Anchor Synthesis (cPAS). The library construction method and sequencing process are carried out according to the following steps:   1. Take appropriate amount of total RNA samples, and add oligo-dT reverse transcription primer and denature the total RNA sample by heat;  2. Add the reverse mix reagent, and reverse transcribed to first-strand cDNA by SMART amplification technology;  3. Synthesize the second-strand cDNA，and use magnetic beads to purify the cDNA, and validate the cDNA;  4. The qualified double stranded cDNA，were used to construct the library with transposase;  5. The library was qualified by the Agilent Technologies 2100 bioanalyzer;  6. The library was circularized;  7. Sequencing: The library was amplified to make DNA nanoball (DNB) and sequenced on DNBSEQ platform. DNBSEQ Low Input Single Cell Smart-Seq Eukaryotic mRNA library