Fig. 4: Characteristics of Smart-seq3xpress single-cell libraries after long-term storage in SEQURNA lysis buffer.
From: Introducing synthetic thermostable RNase inhibitors to single-cell RNA-seq
a Bioanalyzer traces of Tn5-tagmented and amplified libraries of HEK293FT single-cell cDNA varying SEQURNA concentration in the tagmentation reaction. X-axis ticks correspond to 35, 100, 300, 500, 1000, 3000, and 10380 base pairs. b Quantified library yield of libraries in (a). Data shown as mean ± standard error of n = 9 per condition (72 total). c Box plot of number of genes detected in Smart-seq3xpress (SS3xpress) libraries of HEK293FT cells collected in SS3xpress lysis buffer containing no inhibitor (0 U/μl), SEQURNA (0.06, 0.15, 0.3, 0.6, 1.5, or 3.0 U/μL), or standard SS3xpress recombinant RNase inhibitor (RRI). n = 88–96 cells per condition, total = 757 (Day0 only). d Stacked bar plot of reads mapping to exonic (orange), intronic (blue), or intergenic (green) regions for single-cell HEK293FT SS3xpress corresponding to cells in (c). Data shown as mean ± standard error. e Box plots of number of unique molecular identifiers (UMIs) corresponding to cells in (c). f Box plot of number of genes detected in single-cell HEK293FT SS3xpress varying RNase inhibitor conditions in the lysis buffer and storing cells for 0, 1, 4, or 7 days at 25 °C. n = 57–93 cells per condition for Day 0 and n = 6–48 cells per condition for Days 1–7, total = 1220. g Box plot of number of UMIs detected corresponding to cells in (f). h Box plot of number of genes detected in single-cell HEK293FT SS3xpress varying RNase inhibitor conditions in the lysis buffer and storing cells for 0, 1, 4, 7, or 14 days at 4 °C. n = 57–96 cells per condition for Day0 and n = 11–48 cells per condition for Days 1–7, total = 1448. i Box plot of number of UMIs detected corresponding to the cells in (h). c, e–i Data shown as median, interquartile range (IQR), and 1.5x IQR. f–i Data downsampled to 100,000 reads per cell (3456 cells were sequenced pre-downsampling) and red lines indicate medians for RRI. Data underlying relevant plots is provided in Source Data and P-values for statistical comparisons are available in Supplementary Data 1.
