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Low-cost, low-input RNA-seq protocols perform nearly as well as high-input protocols
Peter A. Combs1 and Michael B. Eisen2,3
1 Graduate Program in Biophysics, University of California, Berkeley, California, United States of America,
2 Department of Molecular and Cell Biology, University of California, Berkeley, California, United States of America,
3 Howard Hughes Medical Institute, University of California, Berkeley, California, United States of America,
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.
Data