Fig. 2: Performance of BOLT-seq. | Experimental & Molecular Medicine

Fig. 2: Performance of BOLT-seq.

From: Cost and time-efficient construction of a 3′-end mRNA library from unpurified bulk RNA in a single tube

Fig. 2

Reproducibility of the BOLT-seq method All experimental replicates were subsampled to 1 M. Venn diagrams in (a) represent overlapping subsets of genes detected in each of three replicate BOLT-seq samples using 1000 HEK293T (left) or A549 (right) cells per well in a 96-well microtiter plate. b Normalized gene read counts from each of the three replicate experiments per cell line shown in (a) were compared pairwise. Correlation values (r) for each pairwise comparison are shown in the upper left corner of each panel. Upper panels, HEK293T cells; lower panels, A549 cells. c Correlation between unique normalized ERCC read counts in two experimental BOLT-seq replicates using 1000 HEK293T cells. Left panel, NEBNext; right panel, BOLT-seq. d Correlation between observed and expected ERCC read counts using NEBNext (left) or BOLT-seq (right). e The relationship between the ERCC concentration and probe length using NEBNext (left), BOLT-seq 1/30 (middle), and BOLT-seq 1/100 (right). Undetected dropouts are indicated by a black circle.

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