Data analysis
Data analysis for the TempO-Seq™ assay is fast, accurate, and simple. Our ligated 50-mer Detector Oligos are designed to maximize the distance between all probes making each sequence unique. This design greatly simplifies the data analysis portion because reads produced by the assay are unambiguously assigned to a specific 50-mer associated with a gene, reducing alignment to a lookup table. Using our TempO-SeqR analysis software package, reads generated by TempO-Seq can be aligned to each targeted gene and a table of gene identity vs abundance quickly generated using a standard laptop computer.
Figure 4. Principle component analysis of expression profiles.
TempO-Seq accurately maps > 95% of HTS reads with little or no false positives (reads mapping to the wrong genes). TempO-Seq is sequencing platform independent and available as an easy to install standalone package that is able to align reads, output standard quality control metrics for HTS data, normalize the data and calculate average and standard deviation for each gene between replicate measurements, and provide several different types of data visualization including heat maps (Figure 3) or PCA (Figure 4) for data analysis. Future versions of TempO-Seq will include differential expression analysis routines, pathway analysis, and higher dimensions of data visualization.