I cannot obtain summary alignment with Bowtie2
1
0
Entering edit mode
12 months ago

Hello everyone,

I am aligning my Small RNA Seq data with Bowtie 2. Although the alignment performs well, the only information I obtain after finishing running the alignment is the following:

       ## Time loading reference: 00:00:00
       ## Time loading forward index: 00:00:00
       ## Time loading mirror index: 00:00:00
       ## Multiseed full-index search: 00:00:00
       ## Time searching: 00:00:00
       ## Overall time: 00:00:00
       ## Time loading reference: 00:00:00
       ## Time loading forward index: 00:00:00
       ## Time loading mirror index: 00:00:00
       ## Multiseed full-index search: 00:00:01
       ## Time searching: 00:00:01
       ## Overall time: 00:00:01
       ## Time loading reference: 00:00:00
       ## Time loading forward index: 00:00:00
       ## Time loading mirror index: 00:00:00
       ## Multiseed full-index search: 00:00:00
       ## Time searching: 00:00:00
       ## Overall time: 00:00:00
       ## Time loading reference: 00:00:00
       ## Time loading forward index: 00:00:00
       ## Time loading mirror index: 00:00:00
       ## Multiseed full-index search: 00:00:00
       ## Time searching: 00:00:00
       ## Overall time: 00:00:00
       ## Time loading reference: 00:00:00
       ## Time loading forward index: 00:00:00
       ## Time loading mirror index: 00:00:00
       ## Multiseed full-index search: 00:00:00
       ## Time searching: 00:00:00
       ## Overall time: 00:00:00
       ## Time loading reference: 00:00:00
       ## Time loading forward index: 00:00:00
       ## Time loading mirror index: 00:00:00
       ## Multiseed full-index search: 00:00:00
       ## Time searching: 00:00:00
       ## Overall time: 00:00:00

This is my command line to build the index:

bowtie2-build -o 3 -t 5 Nuevofastaorina human_mirs

And this is my command line to do the alignment:

bowtie2 -x human_mirs -U 1.lane.clean.gz -N 1 --norc --no-unal --no-head --ignore-quals –-quiet > sr1prueba.sam

Does anyone know how to obtain an alignment summary instead of the above information? What I mean by alignment summary is the following:

20000 reads; of these:
  20000 (100.00%) were unpaired; of these:
    1247 (6.24%) aligned 0 times
    18739 (93.69%) aligned exactly 1 time
    14 (0.07%) aligned >1 times
93.77% overall alignment rate

Thank you in advance for your help!

Alejandra

Bowtie2 alignmentsummary SmallRNASeq • 795 views
ADD COMMENT
0
Entering edit mode

I confess I don't know bowtie well, but what happens if you remove --quiet?

ADD REPLY
0
Entering edit mode

Logically we should see the alignment stats. Though I am not sure why it is printing those 00:00:00's.

--quiet            Print nothing besides alignments and serious errors.

Nice catch.

ADD REPLY
0
Entering edit mode
12 months ago
Barry Digby ★ 1.3k

I know the below works for bowtie, you need to capture the stderr. Let me know if it works!

(bowtie2 -x human_mirs -U 1.lane.clean.gz -N 1 --norc --no-unal --no-head --ignore-quals –-quiet > sr1prueba.sam) 2> output.err
ADD COMMENT
0
Entering edit mode

If they are not capturing stderr then where are those 00:00:00 messages coming from I wonder?

ADD REPLY
0
Entering edit mode

They must have carried over the parameter -t/--ftabchars <int> # of chars consumed in initial lookup (default: 10) in the bowtie2-build command to the alignment command.

For bowtie2 it corresponds to: -t/--time print wall-clock time taken by search phases.

Doesn't explain why it was printed 6/7 times though ;) perhaps a for loop we are not privy to

ADD REPLY

Login before adding your answer.

Traffic: 889 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6