# DESeq2 or EdgeR or Limma without replication

**URL:** https://help.galaxyproject.org/t/deseq2-or-edger-or-limma-without-replication/2802
**Category:** Resources
**Tags:** limma\_voom, resources, transcriptomics, deseq2, edger
**Created:** [January 14, 2020, 1:25am UTC](https://help.galaxyproject.org/t/deseq2-or-edger-or-limma-without-replication/2802 "2020-01-14T01:25:33Z")
**Posts on this page:** 1
**Showing post:** 7

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### Author: ![jennaj](https://sea2.discourse-cdn.com/flex020/user_avatar/help.galaxyproject.org/jennaj/32/27_2.png) [@jennaj](https://help.galaxyproject.org/u/jennaj)
#### Post date: [April 15, 2020, 6:55pm UTC](https://help.galaxyproject.org/t/deseq2-or-edger-or-limma-without-replication/2802/7 "2020-04-15T18:55:00Z")

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Replicates will always be required when running these tools in Galaxy. If you really need to do exploratory non-replicate analysis, then using the tools line-command is the standard alternative.

Apologies – but this requirement in the Galaxy-wrapped version of the tools was necessary to avoid wide-spread odd job errors (that were not only confusing to troubleshoot but draining shared public computational resources, often for no practical purpose) and _even more importantly_, to alert scientists about problematic experimental design even if/when putatively “successful” green results were produced.

We tried not requiring replicates originally, but very quickly realized it wasn’t working out well for the vast majority of scientists.

> [@jennaj](#):
>
> FAQ: [Extended Help for Differential Expression Analysis Tools](https://galaxyproject.org/support/diff-expression/)
> 
> The FAQ includes a link to the Bioconductor forum where there is _much_ discussion about their strong recommendations about use of replicates with their tools. Punch line: not using replicates produces invalid results (…if the job completes at all…).

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