Udaondo Zulema, Sittikankaew Kanchana, Uengwetwanit Tanaporn, Wongsurawat Thidathip, Sonthirod Chutima, Jenjaroenpun Piroon, Pootakham Wirulda, Karoonuthaisiri Nitsara, Nookaew Intawat
Department of Biomedical Informatics, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathum Thani 12120, Thailand.
Life (Basel). 2021 Aug 23;11(8):862. doi: 10.3390/life11080862.
With the advantages that long-read sequencing platforms such as Pacific Biosciences (Menlo Park, CA, USA) (PacBio) and Oxford Nanopore Technologies (Oxford, UK) (ONT) can offer, various research fields such as genomics and transcriptomics can exploit their benefits. Selecting an appropriate sequencing platform is undoubtedly crucial for the success of the research outcome, thus there is a need to compare these long-read sequencing platforms and evaluate them for specific research questions. This study aims to compare the performance of PacBio and ONT platforms for transcriptomic analysis by utilizing transcriptome data from three different tissues (hepatopancreas, intestine, and gonads) of the juvenile black tiger shrimp, . We compared three important features: (i) main characteristics of the sequencing libraries and their alignment with the reference genome, (ii) transcript assembly features and isoform identification, and (iii) correlation of the quantification of gene expression levels for both platforms. Our analyses suggest that read-length bias and differences in sequencing throughput are highly influential factors when using long reads in transcriptome studies. These comparisons can provide a guideline when designing a transcriptome study utilizing these two long-read sequencing technologies.
凭借美国太平洋生物科学公司(位于加利福尼亚州门洛帕克)的PacBio和英国牛津纳米孔技术公司的ONT等长读长测序平台所具备的优势,基因组学和转录组学等各个研究领域都能够利用它们的优势。选择合适的测序平台对于研究成果的成功无疑至关重要,因此有必要对这些长读长测序平台进行比较,并针对特定的研究问题对它们进行评估。本研究旨在通过利用幼年黑虎虾三种不同组织(肝胰腺、肠道和性腺)的转录组数据,比较PacBio和ONT平台在转录组分析方面的性能。我们比较了三个重要特征:(i)测序文库的主要特征及其与参考基因组的比对,(ii)转录本组装特征和异构体鉴定,以及(iii)两个平台基因表达水平定量的相关性。我们的分析表明,在转录组研究中使用长读长时,读长偏差和测序通量差异是极具影响力的因素。这些比较可为利用这两种长读长测序技术设计转录组研究提供指导。