Validation of methods for Low-volume RNA-seq

1 Recently, a number of protocols extending RNA-sequencing to the 2 single-cell regime have been published. However, we were concerned that 3 the additional steps to deal with such minute quantities of input sam4 ple would introduce serious biases that would make analysis of the data 5 using existing approaches invalid. In this study, we performed a critical 6 evaluation of several of these low-volume RNA-seq protocols, and found 7 that they performed slightly less well in metrics of interest to us than a 8 more standard protocol, but with at least two orders of magnitude less 9 sample required. We also explored a simple modification to one of these 10 protocols that, for many samples, reduced the cost of library preparation 11 to approximately $20/sample. 12