RNAseq of Thesium chinense in different periods
In this project,  Oligo(dT) magnetic beads were used to enrich the polyA-tailed mRNA from total RNA.    The first-strand cDNA was  synthesized using a 6-base random primer and reverse transcriptase,  with the RNA as the template.    The second-strand cDNA was then synthesized using the first-strand cDNA as the  template.  After the library was constructed, the Qubit2.0 Fluorometer was used for preliminary quantification, the library was diluted to 1.5ng/ul, and then the Agilent 2100 bioanalyzer was used to detect the insert size of the library. The effective concentration of the library was accurately quantified by qRT-PCR (the effective concentration of the library was higher than 2nM) to ensure the quality of the library. After library inspection, different libraries were pooled according to the requirements of effective concentration and target amount of data, and then Illumina sequencing was performed. The basic principle of Sequencing is Sequencing by Synthesis. Four kinds of fluorescently labeled dNTP, DNA polymerase and adaptor primers were added to the sequencing flow cell for amplification. When extending complementary strand in each sequencing cluster, the corresponding fluorescence could be released by adding a fluorescently labeled dNTP. The sequencer captured the fluorescence signal and converted the light signal into sequencing peaks by computer software. Thus, the sequence information of the tested fragment was obtained.