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Creating Graphs

IGB creates these can create several useful graphs from a selected trackannotation tracks. Although the first two graphs (Coverage and Depth) Depth graph can be utilized for all annotations, their the usefulness shines, in combination with the third Mismatch graph (Mismatch) , with .bam and other short read alignment files. Here, you can see where reads are (Coverage), how many reads are present (depth graphat each nucleotide position (Depth) and where mismatches occur as compared to a the genomic sequence file.

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Depth Graph

You can generate an overview of the relative density of annotation coverage across an entire chromosome.  The Make Annotation Coverage Track Depth Graph feature dynamically graphs the relative number of annotations in a track that are present at each pixel across the main window.  As you zoom in and each pixel represents a smaller number of nucleotides, the graph automatically adjusts to indicate the relative number of annotations present in the smaller region covered by that pixel.  nucleotide across the genomic sequence. When you are completely zoomed in, and each pixel represents a single nucleotide base, the graph shows either a 0 (no annotation is present) or a 1 (an annotation is present) for each pixel.To show the density of annotation coverage for an annotation type that you are viewing:actual values for the number of reads aligned at that genomic location. Using the Select tool to hover over a particular point in the graph will open a tooltip with the actual number of reads at that point.

Right-click on the annotation

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in the

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label panel and choose Make Annotation

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Depth Graph.

The following illustration shows the relative annotation coverage at each pixel when the view is zoomed all the way out.  In this example, suppose you conclude that the density of the circled region is significant and you want to examine it more closely.

After you zoom in to display the relative annotation density within the region that is circled in the illustration above, you might decide to look more closely at the region of greater annotation density at the end of the pictured region.

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picture below shows a fully expanded .bam file, showing the difficulty of seeing any details. The second picture shows just the depth graph (.bam track is 'hidden') with the tool tip showing the number of reads aligned onto that specific genomic location. Image Added

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Mismatch Graph

The mismatch graph is specific to short read alignment files, such as .bam.  The graph shows only the number of mismatched nucleotides across all reads at a specific genomic location, which can be very helpful in the detection of allelic variation, SNP identification and also error checking.

This function requires that the genomic sequence be loaded; if it is not loaded at the time you request the graph, IGB will produce a warning with the option to load the sequence for you. If you hover the Select tool over an individual bar of the graph, you will get a tool tip showing the total number of reads, and then the number of reads broken down by nucleotide.

In the picture below, you can see that the total number of reads at the specified position is 260 (blue arrow). Of those reads, 257 at at 'T', which you can see is the matching nucleotide (purple arrows). 3 of those reads contain a 'G' instead; since G is a MISmatch, the graph shows a bar with height of 3 (red arrows). Image Added