Systematic evaluation and optimization of the experimental steps in RNA G-quadruplex structure sequencing
Publication in refereed journal

香港中文大學研究人員
替代計量分析
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其它資訊
摘要cDNA library preparation is important for many high-throughput sequencing applications, such as RNA G-quadruplex structure sequencing (rG4-seq). A systematic evaluation of 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 apply the improved methods to fragmented cellular RNA, and show reduced RNA input requirement, lower transcript abundance variations between biological replicates, as well as lower transcript coverage bias when compared to prior arts. 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.
著者Pui Yan Yeung, Jieyu Zhao, Eugene Yui-Ching Chow, Xi Mou, HuiQi Hong, Leilei Chen, Ting-Fung Chan, Chun Kit Kwok
期刊名稱Scientific Reports
出版年份2019
月份5
日期30
卷號9
出版社Nature Research
文章號碼8091
國際標準期刊號2045-2322
語言美式英語

上次更新時間 2021-08-01 於 23:04