Supplementary Figure 2: Optimization of MARS-seq. | Nature Protocols

Supplementary Figure 2: Optimization of MARS-seq.

From: MARS-seq2.0: an experimental and analytical pipeline for indexed sorting combined with single-cell RNA sequencing

Supplementary Figure 2

(A) Background noise estimation for high and low volume protocol versions. Shown is the average number of UMIs in wells containing single mouse ES (y-axis) vs. an average number of UMIs in negative control wells (x-axis) for two protocol versions with high (green) and low (black) liquid volumes in the RT step. (B) Background noise estimation for different second strand synthesis enzymes mixtures. Shown is the average number of UMIs in common myeloid progenitor cells (CMP; y-axis) vs. an average number of UMIs in negative controls wells (x-axis). Second strand synthesis was performed with ‘A’ composition (green symbols), with addition of RNaseH or without DNA polymerase, or with ‘B’ composition (black symbols) in dilutions of 1:4 or 1:8. Dashed lines demarcate background noise levels of 5% (red) and 10% (magenta).

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