I have both and don't know what to do with either (i.e. how to analyze). I was going to use NCBI genome workbench since it seemed to be the least convoluted and it works with windows.
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Sorted BAM has the data sorted by chromosomes/contigs/scaffolds whatever is in your reference genome. In order to efficiently display/access the data the BAM file has to be sorted.
For viewing aligned/sorted BAM files use integrated genome viewer (IGV) on windows. Unfortunately Broad's website appears to be down for maintenance till Monday but the URL you will want is: http://www.broadinstitute.org/igv/
You can try IGB in the mean time: http://bioviz.org/igb/index.htmlLast edited by GenoMax; 10-17-2013, 02:17 PM.
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Thanks for the info. Unfortunately, it's IGB is not allowing me to install the program and keeps saying error:could not create a virtual java machine.Originally posted by GenoMax View PostSorted BAM has the data sorted by chromosomes/contigs/scaffolds whatever is in your reference genome. In order to efficiently display/access the data the BAM file has to be sorted.
For viewing aligned/sorted BAM files use integrated genome viewer (IGV) on windows. Unfortunately Broad's website appears to be down for maintenance till Monday but the URL you will want is: http://www.broadinstitute.org/igv/
You can try IGB in the mean time: http://bioviz.org/igb/index.html
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I used Bowtie 2 to align against MM10 (mouse genome).Originally posted by GenoMax View PostWhat genome are you working with?
Have you indexed your sorted BAM?Here is another option to try: http://bioinf.scri.ac.uk/tablet/Code:$samtools index sorted.bam
I have not yet created an indexed BAM since the bowtie 2 manual didn't mention anything about that. But it looks like it's required, so I"ll do that tomorrow when I'm at my Linux computer.
It looks like my SAM file is aligning to IGB mRNA but I don't know what the genes are since they're given cryptic names.
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You can download the mm10 GTF file here: ftp://ftp.ensembl.org/pub/current_gt...Cm38.73.gtf.gz IGB should be able to open that and give you gene annotations.Originally posted by KnowNothing2 View PostI used Bowtie 2 to align against MM10 (mouse genome).
It looks like my SAM file is aligning to IGB mRNA but I don't know what the genes are since they're given cryptic names.
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Okay, I figured if I just downloaded an MM10 genome I'd be able to reference (right now I'm using Tablet). that said, I began downloading the mm10 genome from the Bowtie 2 website. Hopefully that does it.Originally posted by GenoMax View PostYou can download the mm10 GTF file here: ftp://ftp.ensembl.org/pub/current_gt...Cm38.73.gtf.gz IGB should be able to open that and give you gene annotations.
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I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.
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Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.
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