# General information on Ribo Zero Plus Microbiome

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

* This product contains 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).

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

* 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).
* 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, rather than 2 x 75 bp, for better analysis results.

**How to analyze Ribo-Zero Plus Microbiome sequencing data?**

* One option it to use the Microbiome Metatranscriptomics App In BaseSpace Sequence Hub.
  * This app is optimized for data prepared using the Illumina Stranded Total RNA with Ligation, Ribo-Zero Plus Microbiome.

**See also:**

* 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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