High-throughput and quantitative genome-wide messenger RNA sequencing for molecular phenotyping
High-throughput and quantitative genome-wide messenger RNA sequencing for molecular phenotyping
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DOI：
10.1186/s12864-015-1788-6
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发表时间：
2015-08-05
期刊：
影响因子：
4.4
通讯作者：
Busch-Nentwich, Elisabeth M.
中科院分区：
文献类型：
--
作者：
Collins, John E.;Wali, Neha;Busch-Nentwich, Elisabeth M.
背景：我们提出了一种全基因组信使RNA(信使RNA)测序技术，将许多样本中的少量RNA转换为分子表型。它涵盖了从样品制备到序列分析的所有步骤，适用于基线分析或扰动测量。结果：转录本3‘端的多重测序识别出不依赖于基因注释的差异转录本丰度。我们发现，在保持测序总量不变的情况下，增加生物重复数可以识别出更丰富的差异转录。结论：该方法可以在任何具有注释的参考基因组的生物体的多腺化RNA上实施，也可以在任何可以获得Illumina测序的实验室中实施。
Background: We present a genome-wide messenger RNA (mRNA) sequencing technique that converts small amounts of RNA from many samples into molecular phenotypes. It encompasses all steps from sample preparation to sequence analysis and is applicable to baseline profiling or perturbation measurements.Results: Multiplex sequencing of transcript 3' ends identifies differential transcript abundance independent of gene annotation. We show that increasing biological replicate number while maintaining the total amount of sequencing identifies more differentially abundant transcripts.Conclusions: This method can be implemented on polyadenylated RNA from any organism with an annotated reference genome and in any laboratory with access to Illumina sequencing.