Additional file 15: Figure S11. of Increasing quality, throughput and speed of sample preparation for strand-specific messenger RNA sequencing

Total RNA input titration using the intermediate ssRNA-seq pipeline. The protocol evaluated includes all changes (1st strand cDNA synthesis, optimal bead purifications, new library construction chemistry with modified ligation condition, bead-based size selection, and UNG treatment) with the exception of the mRNA isolation improvements. (A) Comparable mapping of reads to the human genome. (B) Comparable mapping of reads to various transcriptome catagories. (C) High correlation of expression between lower input and higher input libraries. Pearson’s correlation coefficient was calculated pair-wise as indicated. Heat map was generated for the resulting values with color intensity representing the degree of correlation as per the depicted legend. n = 3 for all inputs except for 2μg and 5μg input amounts where only duplicates were represented. Error bars = Standard Deviation. (JPEG 27 kb)