低成本、低投入的 RNA-seq 方案与高投入方案的性能几乎相当。
Low-cost, low-input RNA-seq protocols perform nearly as well as high-input protocols.
机构信息
Graduate Program in Biophysics, University of California , Berkeley, CA , USA.
Department of Molecular and Cell Biology, University of California , Berkeley, CA , USA ; Howard Hughes Medical Institute, University of California , Berkeley, CA , USA.
出版信息
PeerJ. 2015 Mar 26;3:e869. doi: 10.7717/peerj.869. eCollection 2015.
Recently, a number of protocols extending RNA-sequencing to the single-cell regime have been published. However, we were concerned that the additional steps to deal with such minute quantities of input sample would introduce serious biases that would make analysis of the data using existing approaches invalid. In this study, we performed a critical evaluation of several of these low-volume RNA-seq protocols, and found that they performed slightly less well in per-gene linearity of response, but with at least two orders of magnitude less sample required. We also explored a simple modification to one of these protocols that, for many samples, reduced the cost of library preparation to approximately $20/sample.
最近，已经有许多将 RNA 测序扩展到单细胞水平的方案被公布。然而，我们担心处理如此微量输入样本的额外步骤会引入严重的偏差，从而使使用现有方法分析数据变得无效。在这项研究中，我们对其中的几个低容量 RNA 测序方案进行了严格评估，发现它们在基因响应的线性度方面表现稍差，但所需样本量至少少了两个数量级。我们还探索了对其中一个方案的简单修改，对于许多样本来说，该修改将文库制备的成本降低到每个样本约 20 美元。
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