the size of bam file is about 26 Gb, so I convert it to bedgraph format with Homer, and then to TDF file that would be open easy with IGV, then you mean I can't use this file to see small regions!
Hello,
I want to ask you what is the best way for visualizing big file (Bam), I am going to sort bam file with samtools sort at first, then convert it to bedgraph by this command: bedtools genomecov -ibam *.bam -bg -scale 10.0, and visualize it by IGV. this way is right and certain in your idea or what else?
thanks in advance
2 answers
If you want to be able to see content regardless of the zoom level then make a bigWig files (e.g., with bamCoverage from deepTools or from the bedGraph file bedtools genomecov produces). But if you're going to zoom in and look at smaller regions (<20kb) anyway then just stick with the indexed and sorted BAM file. BAM will still be slower to render than bigWig data, but it otherwise works fine.
If I meant that you couldn't use that to see small regions I would have written that. TDF is an IGV-specific format, as opposed to bigWig files which can be used in other programs, thus making them generally preferable.
You can direcly open your sorted and indexed bam file in IGV. It's not required to convert it to bedgraph.
but is too big and IGV can't open it and will crush!
What is the RAM of your computer ? What is the size of you bam file ? Which type of data ? If DNA-Seq (WGS, WES, ..) what will be the coverage, etc...
Is it imperative to convert to bigWig in case of ChIPseq data?
If that's relevant, let's assume I'm interested in several specific loci/genes
IGV doesn't load the whole bam into the memory, only the section being active viewed. For bam files, IGV will display reads only after you zoom considerably into a small chromosomal region.
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What is the aim of your visualisation? Are you specifically interested in a single gene or locus?