Next week is the AGBT genome conference in Marco Island, Florida. At the conference we will present a poster on work we have been doing with Next Gen Sequencing data analysis. In this post we present the abstract. We'll post the poster when we return from sunny Florida.
The volumes of data that can be obtained from Next Generation DNA sequencing instruments make several new kinds of experiments possible and new questions amenable to study. The scale of subsequent analyses, however, presents a new kind of challenge. How do we get from a collection of several million short sequences of bases to genome-scale results? This process involves three stages of analysis that can be described as primary, secondary, and tertiary data analyses. At the first stage, primary data analysis, image data are converted to sequence data. In the middle stage, secondary data analysis, sequences are aligned to reference data to create application-specific data sets for each sample. In the final stage, tertiary data analysis, the data sets are compared to create experiment-specific results. Currently, the software for the primary analyses is provided by the instrument manufacturers and handled within the instrument itself, and when it comes to the tertiary analyses, many good tools already exist. However, between the primary and tertiary analyses lies a gap.
In RNA-Seq, the process of determining relative gene expression means that sequence data from multiple samples must go through the entire process of primary, secondary, and tertiary analysis. To do this work, researchers must puzzle through a diverse collection of early version algorithms that are combined into complicated workflows with steps producing complicated file formats. Command line tools such as MAQ, SOAP, MapReads, and BWA, have specialized requirements for formatted input and output and leave researchers with large data files that still require additional processing and formatting for tertiary analyses. Moreover, once reads are aligned, datasets need to be visualized and further refined for additional comparative analysis. We present a solution to these challenges that closes the gaps between primary, secondary, and tertiary analysis by showing results from a complete workflow system that includes data collection, processing and analysis for RNA-seq.
And, if you cannot be in sunny Florida, join us in Memphis where we will help kick off the ABRF conference with a workshop on Next Generation DNA Sequencing. I'm kicking the workshop off with a talk entitled "From Reads to Data Sets, Why Next Gen is Not Like Sanger Sequencing."