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Low-cost, low-input RNA-seq protocols perform nearly as well as high-input protocols.

ABSTRACT: 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.

SUBMITTER: Combs PA 

PROVIDER: S-EPMC4380159 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Low-cost, low-input RNA-seq protocols perform nearly as well as high-input protocols.

Combs Peter A PA   Eisen Michael B MB  

PeerJ 20150326

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  ...[more]

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