> For the complete documentation index, see [llms.txt](https://knowledge.illumina.com/llms.txt). Markdown versions of documentation pages are available by appending `.md` to page URLs; this page is available as [Markdown](https://knowledge.illumina.com/library-preparation/rna-library-prep/library-preparation-rna-library-prep-faq-list/000006974.md).

# General information on Ribo Zero Plus Microbiome

**What is Ribo-Zero Plus Microbiome?**

* This product contains the [Ribo-Zero Plus](https://www.illumina.com/products/by-type/molecular-biology-reagents/ribo-zero-plus-rrna-depletion.html) rRNA removal probes and additional rRNA depletion probes designed to target species found in the microbiome of human adult and infant stool samples.
* Ribo-Zero Plus Microbiome can either be purchased as a standalone product, or bundled with library prep reagents in the Illumina Stranded Total RNA Prep Ligation with Ribo-Zero Plus Microbiome kit, listed on the product page [here](https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/stranded-total-rna-prep.html). The standalone product can also be paired with [Illumina Rapid Total RNA Prep.](https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/rapid-total-rna-prep.html)

**Which other sample types are compatible with Ribo-Zero Plus Microbiome?**

* Illumina has reports of effective depletion from skin, vaginal, and oral samples in addition to stool samples, though results may vary.

**How much input is needed? What quality does it need to be?**

* The Illumina Stranded Total and Illumina Rapid Total RNA Prep with Ribo-Zero Plus Microbiome protocols are optimized for 25-500 ng of total RNA input.
* RNA is recommended to have a RIN of at least 5.

**How to use the Ribo-Zero Plus Microbiome reagents?**

* For Illumina Stranded Total RNA Prep Ligation with Ribo-Zero Plus Microbiome, the full protocol is in the updated [Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus, Ribo-Zero Plus Microbiome Reference Guide](https://support.illumina.com/downloads/illumina-stranded-total-rna-reference-1000000124514.html) (1000000124514 v03 or later).
* For Illumina Rapid Total RNA Prep with Ribo-Zero Plus Microbiome, the full protocol is in the Illumina Rapid Total RNA Prep with Ribo-Zero Plus Product Documentation (200077002 v00 or later).
* Briefly, DPM (Microbiome Depletion Probe Pool) is added to DP1 (Ribo-Zero Plus Depletion Probe Pool) to deplete all rRNA species in a single step.

**What are the sequencing recommendations for Ribo-Zero Plus Microbiome?**

* Illumina recommends a 2 x 150 bp read length for Illumina Stranded Total RNA Prep Ligation with Ribo-Zero Plus Microbiome.
* Illumina recommends a read length of 2 x 50 bp or 1 x 100bp for Gene Expression on 100 cycle kits, 2 x 100bp for Variant detection on 200 cycle kits. For additional details, see [Read Depth Requirements for Illumina Rapid RNA kits.](https://github.com/illumina-swi/illumina-knowledge/tree/master/articles/Knowledge/Read-Depth-Requirements-for-Illumina-Rapid-RNA-kits/README.md)

**For more information see**:

* Ribo-Zero Plus Microbiome Depletion Kit [data sheet](https://www.illumina.com/content/dam/illumina/gcs/assembled-assets/marketing-literature/ribo-zero-plus-microbiome-data-sheet-m-gl-00916/ribo-zero-microbiome-data-sheet-m-gl-00916.pdf).
* Ribosomal RNA depletion for metatranscriptomic studies [app note](https://www.illumina.com/content/dam/illumina/gcs/assembled-assets/marketing-literature/ribo-zero-plus-microbiome-app-note-m-gl-00692/ribozero-microbiome-app-note-m-gl-00692.pdf).

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| *For any feedback or questions regarding this article (Illumina Knowledge Article #6974), contact Illumina Technical Support* [*techsupport@illumina.com*](mailto:techsupport@illumina.com?subject=Question%2FFeedback%20Regarding%20Illumina%20Knowledge%20Article%20#000010216%20-%20Library%20Preparation%20\&body=Dear%20Illumina%20Technical%20Support,%0D%0A%0D%0A)*.* |


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