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+# Cow/calf Rumen Metagenomics Pipeline
+
+An end-to-end script to convert Illumina shotgun sequences and metadata into full-blown diversity tables and visualizations. Of course, it's focused on the rumen and dam/calf relationships, but is widely applicable to other systems.
+
+Written entirely during Spring Semester 2019 for work done in [Dr. Hannah Cunningham-Hollinger's lab][hollinger-lab] at the University of Wyoming, computed on UW's [ARCC High-performance servers][arcc-servers] and presented as a [poster] at the Western Section American Association of Animal Science annual meeting.
+
+## Prerequisites
+
+You will need access to the following commands/programs:
+
+- `metaxa2`, `metaxa2_ttt`, `metaxa2_dc` ([Metaxa2])
+- `Rscript` ([R])
+- `source activate` ([Miniconda])
+- `qiime`, `biom` (Install within [conda environment] named `qiime2`)
+
+If working on a HPC, contact your department to find out how to get access to these commands.
+
+## Usage
+
+Clone the script files
+
+```bash
+git clone https://github.com/MillironX/cowcalf-rumen-metagenomic-pipeline.git
+```
+
+Create a directory with all forward- and reverse- read files in it, named as `_R1_001.fastq.gz` for forward-reads and `_R2_001.fastq.gz` for reverse-reads. Add a [QIIME2-compatable metadata file][qiime2-metadata] named `metadata.tsv`, text files containing the minimum and maximum rarefaction values names `rarefaction.min.txt` and `rarefaction.max.txt` and copy all of the code files into it. It should look like
+
+```plaintext
+.
+├── sample1_R1_001.fastq.gz
+├── sample1_R2_001.fastq.gz
+├── sample2_R1_001.fastq.gz
+├── sample2_R2_001.fastq.gz
+├── ...
+├── sampleN_R1_001.fastq.gz
+├── sampleN_R2_001.fastq.gz
+├── metadata.tsv
+├── rarefaction.min.txt
+├── rarefaction.max.txt
+├── main.sh
+├── fastq-to-taxonomy.sh
+├── manipulatefeaturetable.R
+├── fetchmetadata.R
+├── sample-classifier.sh
+└── sample-regression.sh
+```
+
+### With Slurm
+
+These scripts are preconfigured for use with [Slurm] and [Lmod]. Everything is very basic, and should work on any Slurm configuration. Before use, be sure to replace the provided credentials with your own in `main.sh`, `fastq-to-taxonomy.sh`, `sample-classifier.sh`, and `sample-regression.sh`, then run
+
+```bash
+sbatch main.sh
+```
+
+### Without Slurm
+
+Edit `main.sh` and remove every call to `srun` (including its cli options), replace every instance of `$SLURM_NTASKS` with the number of parallel threads you wish to run, and comment out every line that starts `module load`. Then run
+
+```bash
+./main.sh
+```
+
+## Future Work
+
+This project is finished. It is meant to be a reference and an inspiration, but nothing more. I do not intend to update the code now (as embaressing as it might be).
+
+## Known Issues
+
+- Miniconda now uses the `conda activate` command line instead of `source activate`
+
+## License
+
+Distributed under the MIT License. See `LICENSE` for more information.
+
+## Contact
+
+Thomas A. Christensen II - [@MillironX](https://gab.com/MillironX)
+
+Project Link: [https://github.com/MillironX/cowcalf-rumen-metagenomic-pipline](https://github.com/MillironX/cowcalf-rumen-metagenomic-pipline)
+
+[hollinger-lab]: https://www.uwyo.edu/anisci/personnel-directory/wyoming-faculty-and-staff/hannah-cunningham-hollinger/index.html
+[poster]: https://millironx.com/Academia#metagenomics
+[arcc-servers]: https://www.uwyo.edu/arcc/
+[slurm]: https://slurm.schedmd.com/overview.html
+[qiime2-metadata]: https://docs.qiime2.org/2019.4/tutorials/metadata/
+[R]: https://www.r-project.org/
+[metaxa2]: https://microbiology.se/software/metaxa2/
+[Miniconda]: https://conda.io/en/master/miniconda.html
+[conda environment]: https://docs.qiime2.org/2019.4/install/native/#install-qiime-2-within-a-conda-environment
+[Lmod]: https://lmod.readthedocs.io/en/latest/index.html