A cost-effective RNA sequencing protocol for large-scale gene expression studies.

A cost-effective RNA sequencing protocol for large-scale gene expression studies.
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DOI:
10.1038/srep09570
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发表时间:
2015-04-01
期刊:
影响因子:
4.6
通讯作者:
Thomson JA
Thomson JA
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Hou Z;Jiang P;Swanson SA;Elwell AL;Nguyen BK;Bolin JM;Stewart R;Thomson JA

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RNA测序已日益成为生物学研究中不可或缺的工具。虽然近年来测序成本大幅下降,但目前RNA测序的成本仍然是更广泛采用的障碍。在这里,我们提出了一个简单的RNA测序协议,大大降低了成本。该方案使用少至10 ng的总RNA,允许每个泳道多达96个样品的多重测序,并且是链特异性的。 使用人类胚胎干细胞的广泛验证显示了在各种多重水平下技术重复之间的高度一致性。
RNA sequencing has increasingly become an indispensable tool for biological research. While sequencing costs have fallen dramatically in recent years, the current cost of RNA sequencing, nonetheless, remains a barrier to even more widespread adoption. Here, we present a simple RNA sequencing protocol with substantially reduced costs. This protocol uses as little as 10 ng of total RNA, allows multiplex sequencing of up to 96 samples per lane, and is strand specific. Extensive validation using human embryonic stem cells showed high consistency between technical replicates at various multiplexing levels.
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