Department of Equine Industry, Korea National College of Agriculture and Fisheries, Jeonju, 54874, Republic of Korea.
Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for Regenerative Medicine, Dankook University, Cheonan, 31116, Republic of Korea.
Genes Genomics. 2019 Oct;41(10):1233-1240. doi: 10.1007/s13258-019-00853-3. Epub 2019 Jul 26.
The emergence of next-generation sequencing (NGS) technologies has made a tremendous contribution to the deciphering and significance of transcriptome analysis in biological fields. Since the advent of NGS technology in 2007, Illumina, Inc. has provided one of the most widely used sequencing platforms for NGS analysis.
Although reagents and protocols provided by Illumina are adequately performed in transcriptome sequencing, recently, alternative reagents and protocols which are relatively cost effective are accessible. However, the kits derived from various manufacturers have advantages and disadvantages when researchers carry out the transcriptome library construction.
We compared them using a variety of protocols to produce Illumina-compatible libraries based on transcriptome. Three different mRNA sequencing kits were selected for this study: TruSeq RNA Sample Preparation V2 (Illumina, Inc., USA), Universal Plus mRNA-Seq (NuGEN, Ltd., UK), and NEBNext Ultra™ Directional RNA Library Prep Kit for Illumina (New England BioLabs, Ltd., USA). We compared them focusing on cost, experimental time, and data output.
The quality and quantity of sequencing data obtained through the NGS technique were strongly influenced by the type of the sequencing library kits. It suggests that for transcriptome studies, researchers should select a suitable library construction kit according to the goal and resources of experiments.
The present work will help researchers to choose the right sequencing library construction kit for transcriptome analyses.
下一代测序（NGS）技术的出现极大地促进了生物领域转录组分析的破译和意义。自 2007 年 NGS 技术问世以来，Illumina 公司提供了最广泛使用的 NGS 分析测序平台之一。
尽管 Illumina 提供的试剂和方案在转录组测序中得到了充分的应用，但最近，具有相对成本效益的替代试剂和方案也可以获得。然而，当研究人员进行转录组文库构建时，来自不同制造商的试剂盒具有优缺点。
我们使用各种协议进行了比较，基于转录组生成了与 Illumina 兼容的文库。本研究选择了三种不同的 mRNA 测序试剂盒：TruSeq RNA 样品制备 V2（Illumina，Inc.，美国）、Universal Plus mRNA-Seq（NuGEN，Ltd.，英国）和 NEBNext Ultra™ 定向 RNA 文库制备试剂盒用于 Illumina（New England BioLabs，Ltd.，美国）。我们比较了它们的成本、实验时间和数据输出。
通过 NGS 技术获得的测序数据的质量和数量受到测序文库试剂盒类型的强烈影响。这表明，对于转录组研究，研究人员应根据实验的目标和资源选择合适的文库构建试剂盒。
本研究将有助于研究人员为转录组分析选择正确的测序文库构建试剂盒。
