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Updated: Apr 26, 2026
A Converging Strategy for the Generation of a Virtually Sequenced cDNA Library from Unreferenced Pacific Oysters
Published on: June 13, 2019
[Construction and preliminary analysis of a full-length cDNA library for Paris polyphylla var. yunnanensis]
Objective:
A full-length cDNA library of Paris polyphylla var. yunnanensis was constructed in order to research the genes relating to growing development and the genes regulation of its secondary metabolite biosynthesis.
Methods:
The total RNA was extracted from Paris polyphylla var. yunnanensis using modified Trizol method. The SMART (switching mechanism at 5' end of RNA transcript )technology was appliedl to construct the full-length cDNA library. The library titer,recombinant rate and length of insert fragments were determined,the sequences of the library were analyzed by Blastx and were compared to GenBank database.
Results:
The capacity of the library was 2. 5 x 107 cfu/mL, the recombinant rate was 98.5% and the average size of the inserted fragment was 1.5 kb. 9 ESTs (Expressed Sequence Tags) were relating to growing development and 5 ESTs were relating to regulation of secondary metabolite biosynthesis among 149 ESTs obtained from 192 clones sequenced.
Conclusion:
A full-length cDNA library of Paris polyphylla var. yunnanensis is constructed by SMART technology successfully, and the library has enough capacity, high recombinant rate and long insert fragment for the further research to screen and identify the functional genes of Paris polyphylla var. yunnanensis.
