Question: human genome toplevel files
gravatar for biolab
5.1 years ago by
biolab1.2k wrote:

Hi, everyone,

I am new to use human genome files.  I found there are three versions of whole human genome files on Ensembl ( toplevel, hard-masked toplevel, and soft-masked toplevel.  The sizes are quite different. Could anyone please briefly describe to me the differences?

In addition, where is the gff file for download? The above site has a gff file with many regulatory features, e.g., histone methylations. I only need exon, intron, UTRs information.  Thank you very much!

human genome • 3.5k views
ADD COMMENTlink modified 5.1 years ago by Devon Ryan94k • written 5.1 years ago by biolab1.2k
gravatar for Devon Ryan
5.1 years ago by
Devon Ryan94k
Freiburg, Germany
Devon Ryan94k wrote:

The sizes are only different because of file compression. "Masking" refers to manipulating a region in a sequence in some way. Typically, this is done with repeat and low complexity regions so that some aligners (e.g., blast) can avoid them. There are two ways to mask a region in a fasta file. Firstly, one can write its bases in lower case (e.g., "acgt") rather than upper case (e.g., "ACGT"). This is called soft-masking. Secondly, one could instead simply replace repetitive/low complexity regions with an N, termed hard masking. For most cases you'll want to use either the soft-masked or unmasked reference files. If you're using tools like BWA, or tophat or bowtie2 (i.e., almost anything meant to handle NGS data) then the results from using a soft-masked and unmasked reference will be identical (most of these tools simply ignore a base's case). However, should you ever need to use a tool that accounts for masking, then already having a soft-masked genome downloaded can be convenient. For that reason, I personally tend to download the soft-masked versions just so I don't have to bother ever downloading them later.

ADD COMMENTlink written 5.1 years ago by Devon Ryan94k

Hi Devon,

Your explanations are really helpful.  I have just one more question: you mentioned that soft-masked or unmasked genome file should have not effects on mapping (using either tophat2 or bwa), so how's hard masked reference?  What's the side-effects when using the hard-masked file? THANKS a lot!

ADD REPLYlink written 5.1 years ago by biolab1.2k

It's generally a bad idea to use hard-masked files. You're not going to get alignments to stretches of N, so any sequence that you do see that arose from such a region may incorrectly align elsewhere. So using a hard-masked genome is expected to decrease overall mapping quality. The only benefit is that you can map things a bit faster, but that's often a bad trade off.

ADD REPLYlink written 5.1 years ago by Devon Ryan94k

Hi Devon,

Yes, it's bad to use the hard-masked file. Thanks a lot for your detailed answer.

ADD REPLYlink written 5.1 years ago by biolab1.2k
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