Fig. 2: Performance of SEQURNA in single-cell Smart-seq2.
From: Introducing synthetic thermostable RNase inhibitors to single-cell RNA-seq
a Bioanalyzer traces of Smart-seq2 (SS2) cDNA libraries from HEK293FT single cells using varying SEQURNA concentrations (0–24 U/μl) or standard recombinant RNase inhibitor (RRI) in lysis buffer. Tick marks on the x-axis correspond to 35, 100, 300, 500, 1000, 3000, and 10380 base pairs. b Quantified cDNA yield of single-cell SS2 cDNA libraries from HEK293FT cells (fragment size 200–10,000 bp). SEQURNA concentration in lysis buffer on the lower x-axis and resulting concentration in RT on the upper x-axis. Data shown as mean (dot) and standard error (line). n = 3–6 traces per condition (50 traces analyzed). c Percentage primer-dimer of single-cell SS2 cDNA libraries from HEK293FT cells (fragment size 20–50 bp) for the samples in (b). d Box plot of number of genes detected in HEK293FT single-cell SS2 libraries. Data shown as median, interquartile range (IQR), and 1.5x IQR. n = 37–94 per condition (704 libraries analyzed). e Stacked bar plot of fraction reads mapping to exonic (orange), intronic (blue), or intergenic (green) regions of the human genome for single-cell HEK293FT SS2 libraries. Data shown as mean with lines indicating standard error, n = 37–94 per condition. f Line plot of normalized coverage along transcripts for single-cell HEK293FT SS2 libraries, n = 37–94 per condition. g Uniform manifold approximation and projection (UMAP) for single-cell SS2 libraries in mouse cells derived from liver (n = 349 cells) and spleen (n = 368 cells) tissue. Cell sorting was gated for small cell size to enrich cells with low RNA content (Supplementary Fig. 5). Colors indicate tissue origin from liver (blue) or spleen (orange) (left), SEQURNA concentrations 3.0 U/µl (light green), 4.5 U/µl (dark green) or RRI (purple) (middle) and cell type identified as B-cell (blue), T-cell (red), hepatocyte (green) or monocyte (purple) (right). Data underlying relevant plots is provided in Source Data and P-values for statistical comparisons are available in Supplementary Data 1.
