Skip navigation
Run Run Shaw Library City University of Hong KongRun Run Shaw Library

Please use this identifier to cite or link to this item: http://dspace.cityu.edu.hk/handle/2031/9308
Title: Systematic evaluation and optimization of cDNA library preparation for next-generation sequencing
Authors: Yeung, Pui Yan (楊沛欣)
Department: Department of Chemistry
Issue Date: 2019
Course: BCH4036 Project
Programme: Bachelor of Science (Honours) in Applied Biology
Supervisor: Dr. Kwok, Chun Kit
Description: Journal article developed from this OAPS paper: Yeung, P.Y., Zhao, J., Chow, E.Y., Mou, X., Hong, H., Chen, L., ... Kwok, C.K. (2019). Systematic evaluation and optimization of the experimental steps in RNA G-quadruplex structure sequencing. Scientific Reports, 9. doi:10.1038/s41598-019-44541-4
Citation: Yeung, P. Y. (2019). Systematic evaluation and optimization of cDNA library preparation for next-generation sequencing (Outstanding Academic Papers by Students (OAPS), City University of Hong Kong).
Abstract: RNA G-quadruplex (rG4), the secondary structure which forms from stacks of G-quartets in the RNA consists of G-rich sequences, are found to regulate the essential cellular processes in organisms. Moreover, recent research found that rG4 regulates the genes that related to several diseases, including neurological disorder and cancers. Hence, understanding the biology of rG4-containing RNAs might be beneficial to analysis and therapeutic strategy development towards corresponding diseases. cDNA library preparation is important for many high-throughput sequencing applications, such as rG4-seq. A systematic evaluation on the procedures of the experimental pipeline, however, is lacking. Herein, we perform a comprehensive assessment of the 5 key experimental steps involved in the cDNA library preparation of rG4-seq, and identify better reaction conditions and/or enzymes to carry out each of these key steps. Notably, we benchmark on the RNA input requirement for this optimized experimental pipeline, and show that only sub-nanogram level of RNA is required for this new and sensitive approach. In addition, the time to perform these steps is substantially reduced to hours. Our method and results can be directly applied in protocols that require cDNA library preparation, and provide insights to the further development of simple and efficient cDNA library preparation for different biological applications.
Appears in Collections:OAPS - Dept. of Chemistry 

Files in This Item:
File SizeFormat 
fulltext.html156 BHTMLView/Open
Show full item record


Items in Digital CityU Collections are protected by copyright, with all rights reserved, unless otherwise indicated.

Send feedback to Library Systems
Privacy Policy | Copyright | Disclaimer