Pati Pratap Kumar, Ma Liuyin, Hunt Arthur G
Department of Biotechnology, Guru Nanak Dev University, Amritsar, India.
Methods Mol Biol. 2015;1255:159-74. doi: 10.1007/978-1-4939-2175-1_14.
Second generation DNA sequencing technologies have been a great boon for the study of mRNA polyadenylation. The experimental determination of large numbers of polyadenylation sites using high-throughput sequencing strategies has provided the necessary platform for deeper understanding the regulation of gene expression in eukaryotes. For generating large sets of data to map poly-A sites, specialized sample preparations that target the junction of 3'-UTR and the poly(A) tail are usually employed. Here, we describe three different protocols that are effectively used for global determinations of poly(A) site choice in plants.
第二代DNA测序技术极大地促进了对mRNA聚腺苷酸化的研究。利用高通量测序策略对大量聚腺苷酸化位点进行实验测定,为更深入了解真核生物基因表达调控提供了必要的平台。为了生成大量数据来绘制聚腺苷酸位点图谱,通常采用针对3'-UTR与聚(A)尾连接处的专门样本制备方法。在此,我们描述了三种不同的方案,它们有效地用于全面测定植物中聚(A)位点的选择。