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一种用于高效、准确且可重复构建RNA测序文库的自动化方法。

An automated method for efficient, accurate and reproducible construction of RNA-seq libraries.

作者信息

Tsompana Maria, Valiyaparambil Sujith, Bard Jonathan, Marzullo Brandon, Nowak Norma, Buck Michael Joseph

机构信息

Department of Biochemistry and Center of Excellence in Bioinformatics and Life Sciences, State University of New York at Buffalo, 701 Ellicott St., 14203, Buffalo, NY, USA.

出版信息

BMC Res Notes. 2015 Apr 3;8:124. doi: 10.1186/s13104-015-1089-9.

Abstract

BACKGROUND

Integration of RNA-seq expression data with knowledge on chromatin accessibility, histone modifications, DNA methylation, and transcription factor binding has been instrumental for the unveiling of cell-specific local and long-range regulatory patterns, facilitating further investigation on the underlying rules of transcription regulation at an individual and allele-specific level. However, full genome transcriptome characterization has been partially limited by the complexity and increased time-requirements of available RNA-seq library construction protocols.

FINDINGS

Use of the SX-8G IP-Star® Compact System significantly reduces the hands-on time for RNA-seq library synthesis, adenylation, and adaptor ligation providing with high quality RNA-seq libraries tailored for Illumina high-throughput next-generation sequencing. Generated data exhibits high technical reproducibility compared to data from RNA-seq libraries synthesized manually for the same samples. Obtained results are consistent regardless the researcher, day of the experiment, and experimental run.

CONCLUSIONS

Overall, the SX-8G IP-Star® Compact System proves an efficient, fast and reliable tool for the construction of next-generation RNA-seq libraries especially for trancriptome-based annotation of larger genomes.

摘要

背景

将RNA测序表达数据与染色质可及性、组蛋白修饰、DNA甲基化和转录因子结合等知识相结合,有助于揭示细胞特异性的局部和远距离调控模式,促进在个体和等位基因特异性水平上对转录调控潜在规则的进一步研究。然而,全基因组转录组特征分析在一定程度上受到现有RNA测序文库构建方案的复杂性和时间要求增加的限制。

研究结果

使用SX-8G IP-Star® Compact系统可显著减少RNA测序文库合成、腺苷酸化和接头连接的实际操作时间,提供为Illumina高通量下一代测序量身定制的高质量RNA测序文库。与为相同样本手动合成的RNA测序文库的数据相比,生成的数据具有很高的技术重复性。无论研究人员、实验日期和实验运行情况如何,获得的结果都是一致的。

结论

总体而言,SX-8G IP-Star® Compact系统被证明是构建下一代RNA测序文库的高效、快速且可靠的工具,尤其适用于基于转录组的较大基因组注释。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72e2/4391147/e0bed80312f5/13104_2015_1089_Fig1_HTML.jpg

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