Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • Slacanch
    Member
    • Jan 2013
    • 10

    #1

    Hisat2 reporting of uniquely mapped reads

    Recently i've been optimising Hisat2 to work with the latest version of the human genome. According to its manual, Hisat2 reports the number of valid alignments within the tag NH:i:N, where N is the number of valid alignments.

    At the same time, Hisat2 also outputs an alignment summary at the end of a run, here's an example:

    1000000 reads; of these:
    1000000 (100.00%) were unpaired; of these:
    436566 (43.66%) aligned 0 times
    403452 (40.35%) aligned exactly 1 time
    159982 (16.00%) aligned >1 times
    56.34% overall alignment rate


    I decided to isolate uniquely mapping reads from multimappers and based myself on the NH:i tag, here's the code i used to isolate the uniquely mapping reads:

    cat logs/run0000.o7572309 | awk '{if($2 < 256) print $0}' | grep -P '^@|NH:i:1\b'

    here, the awk section gets rid of all secondary alignments, while grep is supposed to isolate all the reads containing the NH:i:1 tags (i.e. uniquely mapping).

    There seems to be a discrepancy between the number of uniquely mapping reads reported at the end of the alignment:

    403452 (40.35%) aligned exactly 1 time

    and the ones that contain the NH:i:1 tag:

    500082


    is there something i'm overlooking? why the discrepancy?

Latest Articles

Collapse

  • kinpict.com
    Reply to 16S/ITS Profiling vs. Shotgun Metagenomics: What Changes in Your Workflow?
    by kinpict.com
    The part about this topic feels surprisingly grounded, and the details on the page is the bit that made me slow down. I’d keep <a href="https://kinpict.com/">ai family photo generator</a> bookmarked for later.
    Today, 06:37 PM
  • SEQadmin2
    16S/ITS Profiling vs. Shotgun Metagenomics: What Changes in Your Workflow?
    by SEQadmin2





    NGS has revolutionized clinical microbiology by offering new ways to identify and surveil pathogens without prior knowledge of the target. One popular approach is 16S rRNA and ITS amplicon sequencing, which targets specific microbial markers. But studies are increasingly incorporating shotgun metagenomics, a technique that sequences all DNA from a sample without targeting specific microbial markers. This approach allows more detailed taxonomic classification,...
    10-30-2026, 06:46 AM

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by SEQadmin2, 11-12-2026, 07:59 AM
0 responses
28 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 11-05-2026, 09:25 AM
0 responses
35 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 11-03-2026, 11:54 AM
2 responses
56 views
0 reactions
Last Post kinpict.com  
Started by SEQadmin2, 10-28-2026, 09:57 AM
0 responses
175 views
0 reactions
Last Post SEQadmin2  
Working...