RNA-seq of RSPO3 overexpression OVCAR3 cell-02
Most mRNAs of eukaryotes had poly(A) structure,and the mRNAs with poly(A) structure were captured by magnetic beads with Oligo(DT). The first strand of cDNA was synthesized in M-MuLV reverse transcriptase system using fragmented mRNA as template and random oligonucleotide as primer. RNA strand was degraded by RNaseH and these condstrand of cDNA was synthesized by dNTPs under the DNA polymerasel system. After terminal repair and joint connection, AMPure XP beads were used to screen about 200bp of cDNA. PCR amplification was carried out and the PCR products were purified by AMPure XP beads again. Finally sequencing libraries were obtained. Illumina PE150 sequencing was performed after quality inspection.