```{r setup, echo=FALSE} stopifnot(BiocInstaller::biocVersion() == "3.0") ``` ```{r style, echo = FALSE, results = 'asis'} BiocStyle::markdown() ``` ## Appendix 1: Installing and using IGV _NOTE_: All instructions in this document should be performed **on your laptop**, not on the RStudio Server AMI. * We'll first create a directory called "igv" in your home directory. You can determine your home directory by issuing the following command in a Terminal window on Linux or Mac: echo $HOME On Windows, open a command window by clicking Start, then Run, then typing `cmd` and pressing Enter. In the window, type echo %USERPROFILE% * So go to the indicated directory, and then issue the command mkdir igv That will create an `igv` directory in your home directory. * Copy the file [hg19_alias.tab](hg19_alias.tab) to this directory. This is a simple tab-delimited file that maps between the sequence names used by the alignment, and the sequence names known to IGV. * Install Java if you haven't already. Go to [https://www.java.com/en/](https://www.java.com/en/) and click on Free Java Download. Download and install. * Download IGV from [https://www.broadinstitute.org/igv/download](https://www.broadinstitute.org/igv/download). There are several ways to do this, the easiest is to click on one of the Java Web Start links. Make sure you have more memory available than what is listed below the Launch button. You may need to change the security settings on your computer to allow IGV to launch. * With IGV running, let's change the default genome. It's currently set to Human hg18, so click on the dropdown in the upper left that says "Human hg18". Click on "More..." and then in the `Filter` box, start typing `hg19`. `Human hg19` will then show up in the box and you can double-click on it. We're now using Human hg19 as our default genome. * You can now open bam files in IGV. Here are the URLs of BAM files for this class. You can open them by clicking on IGV's `File` menu, then clicking on `Load From URL...`. You can paste one of these URLs into the box and click OK: 

    http://s3-us-west-2.amazonaws.com/oct2014bamfiles/SRR1039508_sorted.bam
    http://s3-us-west-2.amazonaws.com/oct2014bamfiles/SRR1039509_sorted.bam
    http://s3-us-west-2.amazonaws.com/oct2014bamfiles/SRR1039512_sorted.bam
    http://s3-us-west-2.amazonaws.com/oct2014bamfiles/SRR1039513_sorted.bam
    http://s3-us-west-2.amazonaws.com/oct2014bamfiles/SRR1039516_sorted.bam
    http://s3-us-west-2.amazonaws.com/oct2014bamfiles/SRR1039517_sorted.bam
    http://s3-us-west-2.amazonaws.com/oct2014bamfiles/SRR1039520_sorted.bam
    http://s3-us-west-2.amazonaws.com/oct2014bamfiles/SRR1039521_sorted.bam
You may see a warning that the bam file does not contain any sequence names which match the current genome. You can ignore this (click OK). * Zoom in to a particular gene, e.g., SPARCL1, by entering the gene symbol in the box toward the top center of the browser window. Then click `Go`. Adjust the zoom until reads come in to view, and interpret the result.