Question: Error with MACS2 peak calling
0
gravatar for 340072256
19 months ago by
340072256 • 0
340072256 • 0 wrote:

Hi dear all, i am new to ChIP-seq analysis and i have been recently using MACS2 callpeaks on galaxy. The but it always fails and i don't know why..... plz help me and thank you all in advance :)

Here's the parameters set

ChIP-Seq Treatment File              42: Chipseq_NSC_H3K4me3_trimmed_unique_BT2.bam 
ChIP-Seq Control File                43: Chipseq_NSC_input_trimmed_unique_BT2.bam   

Are your inputs Paired-end BAM files?   True    
Effective genome size                   2451960000  
Band width for picking regions to compute fragment size 300 

mfold   
Set lower mfold bound   5   
Set upper mfold bound   50  

Peak detection based on qvalue  
Minimum FDR (q-value) cutoff for peak detection              0.05   

Build Model                                                 create_model    

Outputs Peaks as tabular file Peak summits Scores in bedGraph files (--bdg) Summary page (html) Plot in PDF 

Advanced options    off

Here's the error report

Fatal error: Exit code 2 ()

INFO  @ Sat, 29 Apr 2017 13:29:38: 
# Command line: callpeak --name MACS2 -t /galaxy-repl/main/files/019/759/dataset_19759955.dat -c /galaxy-repl/main/files/019/759/dataset_19759956.dat --format BAMPE --gsize 2451960000 --bw 200 --mfold 40 80 --bdg --qvalue 0.05
# ARGUMENTS LIST:
# name = MACS2
# format = BAMPE
# ChIP-seq file = ['/galaxy-repl/main/files/019/759/dataset_19759955.dat']
# control file = ['/galaxy-repl/main/files/019/759/dataset_19759956.dat']
# effective genome size = 2.45e+09
# band width = 300
# model fold = [5, 50]
# qvalue cutoff = 5.00e-02
# Larger dataset will be scaled towards smaller dataset.
# Range for calculating regional lambda is: 1000 bps and 10000 bps
# Broad region calling is off
# Paired-End mode is on

INFO  @ Sat, 29 Apr 2017 13:29:38: #1 read fragment files... 
INFO  @ Sat, 29 Apr 2017 13:29:38: #1 read treatment fragments... 
INFO  @ Sat, 29 Apr 2017 13:29:53:  1000000 
INFO  @ Sat, 29 Apr 2017 13:30:08:  2000000 
INFO  @ Sat, 29 Apr 2017 13:30:23:  3000000 
INFO  @ Sat, 29 Apr 2017 13:30:38:  4000000 
INFO  @ Sat, 29 Apr 2017 13:30:53:  5000000 
INFO  @ Sat, 29 Apr 2017 13:31:08:  6000000 
INFO  @ Sat, 29 Apr 2017 13:31:23:  7000000 
INFO  @ Sat, 29 Apr 2017 13:31:38:  8000000 
INFO  @ Sat, 29 Apr 2017 13:31:53:  9000000 
INFO  @ Sat, 29 Apr 2017 13:32:07:  10000000 
INFO  @ Sat, 29 Apr 2017 13:32:22:  11000000 
INFO  @ Sat, 29 Apr 2017 13:32:37:  12000000 
INFO  @ Sat, 29 Apr 2017 13:32:52:  13000000 
INFO  @ Sat, 29 Apr 2017 13:33:07:  14000000 
INFO  @ Sat, 29 Apr 2017 13:33:20: #1.2 read input fragments... 
INFO  @ Sat, 29 Apr 2017 13:33:36:  1000000 
INFO  @ Sat, 29 Apr 2017 13:33:51:  2000000 
INFO  @ Sat, 29 Apr 2017 13:34:07:  3000000 
INFO  @ Sat, 29 Apr 2017 13:34:22:  4000000 
INFO  @ Sat, 29 Apr 2017 13:34:38:  5000000 
INFO  @ Sat, 29 Apr 2017 13:34:54:  6000000 
INFO  @ Sat, 29 Apr 2017 13:35:10:  7000000 
INFO  @ Sat, 29 Apr 2017 13:35:24: #1 mean fragment size is determined as 221 bp from treatment 
INFO  @ Sat, 29 Apr 2017 13:35:24: #1 note: mean fragment size in control is 216 bp -- value ignored 
INFO  @ Sat, 29 Apr 2017 13:35:24: #1 fragment size = 221 
INFO  @ Sat, 29 Apr 2017 13:35:24: #1  total fragments in treatment: 14330631 
INFO  @ Sat, 29 Apr 2017 13:35:24: #1 user defined the maximum fragments... 
INFO  @ Sat, 29 Apr 2017 13:35:24: #1 filter out redundant fragments by allowing at most 1 identical fragment(s) 
INFO  @ Sat, 29 Apr 2017 13:36:09: #1  fragments after filtering in treatment: 9983930 
INFO  @ Sat, 29 Apr 2017 13:36:09: #1  Redundant rate of treatment: 0.30 
INFO  @ Sat, 29 Apr 2017 13:36:09: #1  total fragments in control: 7630985 
INFO  @ Sat, 29 Apr 2017 13:36:09: #1 user defined the maximum fragments... 
INFO  @ Sat, 29 Apr 2017 13:36:09: #1 filter out redundant fragments by allowing at most 1 identical fragment(s) 
INFO  @ Sat, 29 Apr 2017 13:36:30: #1  fragments after filtering in control: 4959605 
INFO  @ Sat, 29 Apr 2017 13:36:30: #1  Redundant rate of control: 0.35 
INFO  @ Sat, 29 Apr 2017 13:36:30: #1 finished! 
INFO  @ Sat, 29 Apr 2017 13:36:30: #2 Build Peak Model... 
INFO  @ Sat, 29 Apr 2017 13:36:30: #2 Skipped... 
INFO  @ Sat, 29 Apr 2017 13:36:30: #2 Use 221 as fragment length 
INFO  @ Sat, 29 Apr 2017 13:36:30: #3 Call peaks... 
INFO  @ Sat, 29 Apr 2017 13:36:30: #3 Pre-compute pvalue-qvalue table... 
INFO  @ Sat, 29 Apr 2017 13:37:45: #3 In the peak calling step, the following will be performed simultaneously: 
INFO  @ Sat, 29 Apr 2017 13:37:45: #3   Write bedGraph files for treatment pileup (after scaling if necessary)... MACS2_treat_pileup.bdg 
INFO  @ Sat, 29 Apr 2017 13:37:45: #3   Write bedGraph files for control lambda (after scaling if necessary)... MACS2_control_lambda.bdg 
INFO  @ Sat, 29 Apr 2017 13:37:45: #3   Pileup will be based on sequencing depth in control. 
INFO  @ Sat, 29 Apr 2017 13:37:45: #3 Call peaks for each chromosome... 
INFO  @ Sat, 29 Apr 2017 13:40:39: #4 Write output xls file... MACS2_peaks.xls 
INFO  @ Sat, 29 Apr 2017 13:40:39: #4 Write peak in narrowPeak format file... MACS2_peaks.narrowPeak 
INFO  @ Sat, 29 Apr 2017 13:40:40: #4 Write summits bed file... MACS2_summits.bed 
INFO  @ Sat, 29 Apr 2017 13:40:40: Done!
chip-seq • 964 views
ADD COMMENT • link • modified 19 months ago by Devon Ryan • 1.9k • written 19 months ago by 340072256 • 0
1
gravatar for Devon Ryan
19 months ago by
Devon Ryan • 1.9k
Germany
Devon Ryan • 1.9k wrote:

The problem is the PDF output. The "Build Peak Model" step was skipped, so there was nothing to make a PDF out of. This is actually a common problem.

ADD COMMENT • link written 19 months ago by Devon Ryan • 1.9k

Thank you very much ! I have tried to disable the PDF output and it was not failed this time :) But I still don't understand why the " Build peak model" was skipped? isn't MACS2 supposed to call peaks based on the peak model it builds?

ADD REPLY • link written 19 months ago by 340072256 • 0

I'm not sure why, that particular wording should only happen if you instruct MACS2 not to build a model, which is not something you did.

ADD REPLY • link written 19 months ago by Devon Ryan • 1.9k
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