# DiffBind Chip-Seq error **URL:** \ **Category:** Uncategorized\ **Tags:** macs2, chip-seq, epigenetics\ **Created:** [July 13, 2021, 3:02pm UTC](https://help.galaxyproject.org/t/diffbind-chip-seq-error/6358 "2021-07-13T15:02:16Z")\ **Posts on this page:** 6\ **Page:** 1
**Author:** ![dush](https://avatars.discourse-cdn.com/v4/letter/d/a87d85/32.png) [@dush](https://help.galaxyproject.org/u/dush)\ **Post date:** [July 13, 2021, 3:02pm UTC](https://help.galaxyproject.org/t/diffbind-chip-seq-error/6358/1 "2021-07-13T15:02:16Z")
Hi, I have Chipseq data and 2 replicates for each condition. I did MACS2 peak calling and tried to perform DiffBind. But I always get the same error. [1] “en\_US.UTF-8” Can you please help me with this. Thanks ---
**Author:** ![gallardoalba](https://sea2.discourse-cdn.com/flex020/user_avatar/help.galaxyproject.org/gallardoalba/32/1903_2.png) [@gallardoalba](https://help.galaxyproject.org/u/gallardoalba)\ **Post date:** [July 13, 2021, 3:08pm UTC](https://help.galaxyproject.org/t/diffbind-chip-seq-error/6358/2 "2021-07-13T15:08:26Z")
Hi @dush, could you provide additional information about the error report? You can find it in the bug icon: [![](https://img.youtube.com/vi/k8kjcNztQeo/hqdefault.jpg "View output errors") ](https://www.youtube.com/watch?v=k8kjcNztQeo) Regards ---
**Author:** ![dush](https://avatars.discourse-cdn.com/v4/letter/d/a87d85/32.png) [@dush](https://help.galaxyproject.org/u/dush)\ **Post date:** [July 13, 2021, 5:02pm UTC](https://help.galaxyproject.org/t/diffbind-chip-seq-error/6358/3 "2021-07-13T17:02:11Z")
### Detailed Job InformationJob environment and execution information is available at the job [info page](https://usegalaxy.org/datasets/bbd44e69cb8906b563431f779c766fd1/show_params). > Job ID | 36606423 (bbd44e69cb8906b530f67995209c8a3a) | > > - | - | > Tool ID | [toolshed.g2.bx.psu.edu/repos/bgruening/diffbind/diffbind/2.10.0](http://toolshed.g2.bx.psu.edu/repos/bgruening/diffbind/diffbind/2.10.0) | > Tool Version | 2.10.0 | > Job PID or DRM id | 30689932 | > Job Tool Version | R version 3.5.1 (2018-07-02) – “Feather Spray”, DiffBind version 2.10.0, rjson version 0.2.20 | ### Job Execution and Failure Information#### Command Line ```auto ln -s '/galaxy-repl/main/files/057/429/dataset_57429239.dat' 'Treated-MACS2_callpeak_on_T-peaks.bed' && ln -s '/galaxy-repl/main/files/057/429/dataset_57429110.dat' 'Filter_SAM_or_BAM __T-bamreads.bam' && ln -s '/galaxy-repl/main/files/_metadata_files/004/628/metadata_4628948.dat' 'Filter_SAM_or_BAM__ T-bamreads.bai' && ln -s '/galaxy-repl/main/files/057/429/dataset_57429139.dat' 'Filter_SAM_or_BAM __N-bamcontrol.bam' && ln -s '/galaxy-repl/main/files/_metadata_files/004/628/metadata_4628955.dat' 'Filter_SAM_or_BAM__ N-bamcontrol.bai' && ln -s '/galaxy-repl/main/files/057/429/dataset_57429192.dat' 'Control-MACS2_callpeak_control62121_narrow_Peaks-peaks.bed' && ln -s '/galaxy-repl/main/files/057/429/dataset_57429080.dat' 'Filter_SAM_or_BAM __C-bamreads.bam' && ln -s '/galaxy-repl/main/files/_metadata_files/004/628/metadata_4628952.dat' 'Filter_SAM_or_BAM__ C-bamreads.bai' && Rscript '/cvmfs/[main.galaxyproject.org/shed_tools/toolshed.g2.bx.psu.edu/repos/bgruening/diffbind/194e3f2c1d86/diffbind/diffbind.R](http://main.galaxyproject.org/shed_tools/toolshed.g2.bx.psu.edu/repos/bgruening/diffbind/194e3f2c1d86/diffbind/diffbind.R)' -i '[["Condition", [{"Control": ["Filter_SAM_or_BAM__N-bamcontrol.bam"]}, {"Control": ["Filter_SAM_or_BAM__C-bamreads.bam"]}, {"Control": ["Control-MACS2_callpeak_control62121_narrow_Peaks-peaks.bed"]}, {"Treated": ["Filter_SAM_or_BAM__N-bamcontrol.bam"]}, {"Treated": ["Filter_SAM_or_BAM__T-bamreads.bam"]}, {"Treated": ["Treated-MACS2_callpeak_on_T-peaks.bed"]}]]]' -o '/galaxy-repl/main/files/058/461/dataset_58461363.dat' -t 0.5 -f interval -p 'XXXX' -n "8" ``` #### stderr ```auto MACS2_callpeak_control62121_narrow_Peaks MACS2_callpeak_control62121_narrow_Peaks Control NA raw MACS2_callpeak_on_T MACS2_callpeak_on_T Treated NA raw Warning message: No contrasts added. Perhaps try more categories, or lower value for minMembers. Error in pv.DBA(DBA, method, bSubControl, bFullLibrarySize, bTagwise = bTagwise, : Unable to perform analysis: no contrasts specified. Calls: dba.analyze -> pv.DBA Warning message: No contrasts added. Perhaps try more categories, or lower value for minMembers. ``` #### stdout ```auto [1] "en_US.UTF-8" ``` #### Job Information ```auto None ``` #### Job Traceback ```auto None ``` This is an automated message. Do not reply to this address. ---
**Author:** ![gallardoalba](https://sea2.discourse-cdn.com/flex020/user_avatar/help.galaxyproject.org/gallardoalba/32/1903_2.png) [@gallardoalba](https://help.galaxyproject.org/u/gallardoalba)\ **Post date:** [July 13, 2021, 5:42pm UTC](https://help.galaxyproject.org/t/diffbind-chip-seq-error/6358/4 "2021-07-13T17:42:37Z")
Hi @dush, please have a look at this entry; it covers your case and provide some additional info about how to use Diffbind. > [@DiffBind\_ChIP-Seq\_Error\_message](https://help.galaxyproject.org/t/diffbind-chip-seq-error-message/2299/2): > > Hi @shamjdeed At least two replicates per condition are required. And for Diffbind, the first condition must be specifically named “Condition” (quirk in how the underlying wrapped tool works). FAQ: [https://galaxyproject.org/support/](https://galaxyproject.org/support/)[Extended Help for Differential Expression Analysis Tools](https://galaxyproject.org/support/diff-expression/) Regards ---
**Author:** ![dush](https://avatars.discourse-cdn.com/v4/letter/d/a87d85/32.png) [@dush](https://help.galaxyproject.org/u/dush)\ **Post date:** [July 13, 2021, 8:01pm UTC](https://help.galaxyproject.org/t/diffbind-chip-seq-error/6358/5 "2021-07-13T20:01:45Z")
Hi Cristóbal, Thank you so much for the help. I also tried with 2 replicates before and gave the same error. But I didn’t use the name ‘Condition’. I will do that and update you. Thanks again for the help. Best regards Dushani ---
**Author:** ![dush](https://avatars.discourse-cdn.com/v4/letter/d/a87d85/32.png) [@dush](https://help.galaxyproject.org/u/dush)\ **Post date:** [July 15, 2021, 5:51pm UTC](https://help.galaxyproject.org/t/diffbind-chip-seq-error/6358/6 "2021-07-15T17:51:25Z")
Hi Cristóbal, It worked. Thank you! Best regards Dushani