title: RNA-seq of Drosophila melanogaster:leg
design: Genomic RNA was extracted from the leg of fruit fly samples using the TRNzol Universal Reagent kit. The extracted RNA underwent quantification and quality control. According to the operation instructions of the Hieff NGS Ultima Dual-mode RNA Library Prep Kit (12308ES96), different index tags are selected to build the library; For the qualified Total RNA samples, magnetic beads with Oligo(dT) are used to enrich eukaryotic mRNA; Then, Frag/Prime Buffer is added to break the mRNA into short fragments, using mRNA as the template, and synthesizing one strand of cDNA with six-base random primers (random hexamers); Then, dNTPs and Enzyme Master Mix are added to synthesize the second strand of cDNA; After the second strand synthesis, Ligation Enhancer, Novel T4 DNA Ligase and DNA Adapter are added for adapter ligation; Then, DNA Selection Beads are used to purify the double-stranded cDNA; Finally, fragment size selection is performed, and PCR enrichment is carried out to obtain the final cDNA library.