Ginkgo long-chain non-coding RNA and application thereof to bilobalide biosynthesis
A long-chain non-coding, ginkgolide technology, applied in DNA/RNA fragments, applications, recombinant DNA technology, etc., can solve the problem of lncRNA in the blank stage
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[0012] 1. Collection of samples
[0013] The 5-year-old healthy male ginkgo tree was used as the test material, and the leaf and root samples (as a control) were collected respectively, immediately put into liquid nitrogen, and stored at -80°C for future use.
[0014] 2. RNA extraction
[0015] RNA was isolated and purified from Ginkgo biloba leaves and roots using Trizol kit (Invitrogen, Carlsbad, CA, USA). RNA purity and integrity were checked using NanoDrop ND-1000 (NanoDrop, Wilmington, DE, USA) and Agilent 2100, respectively.
[0016] 3. RNA library construction and sequencing
[0017] Via Ribo-Zero TM After removal of rRNA by the rRNA Removal Kit (Illumina, San Diego, USA), the remaining RNA was fragmented into small pieces using divalent cations at high temperature. The cleaved RNA fragments were reverse transcribed into cDNA, which was then used to synthesize U-labeled second-strand DNA using E. coli DNA polymerase I, RNase H, and dUTP. A bases are then added to t...
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