> 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/000001153.md).

# Frequently asked questions regarding the Ribo Zero Plant and Globin kits.

**Note: This article refers to Ribo-Zero, which is a bead-based pull down method of rRNA depletion, used with TruSeq Stranded Total RNA kits. This is different from Ribo-Zero Plus, an enzymatic method of rRNA depletion, used with Illumina Stranded Total RNA kits.**

\*\*Are there any protocol considerations for the Plant or Globin Ribo-Zero kits?\*\*All Ribo-Zero protocols follow the same [TruSeq Stranded Total RNA Reference Guide](https://emea.support.illumina.com/downloads/truseq-stranded-total-rna-reference-guide-1000000040499.html). The only component that differs is the rRNA Removal Mix.

\*\*Do the Ribo-Zero Globin or Plant kits work on degraded RNA?\*\*All Ribo-Zero kits are compatible with degraded RNA. See the Reference Guide for more information.

\*\*Can samples generated with the plant or globin TruSeq protocols be multiplexed in the same lane as human TruSeq RNA samples?\*\*Yes, but the index options are the same across all kits. It is important that all samples are uniquely indexed.

**Ribo-Zero Globin specific questions**

**Which molecules are removed with the Ribo-Zero Globin kit?**\
The Ribo-Zero Globin removal mix targets cytoplasmic rRNA, mitochondrial rRNA, and globin mRNA.

**Does the Ribo-Zero Globin kit subtract Globin mRNA from non-human RNA?**\
The TruSeq Stranded Ribo-Zero Globin kit supports human, mouse, and rat, and may be compatible with other species as well.

**Can the Ribo-Zero Globin kit be used with samples other than blood?**\
The Ribo-Zero Globin kit has been specifically tailored for blood samples from human, mouse, or rat. In addition to removing cytoplasmic and mitochondrial rRNA, it will also deplete globin mRNA.

**Ribo-Zero Plant specific questions**

**Which molecules are removed with the Ribo-Zero Plant kit?**\
The Ribo-Zero Plant removal mix targets cytoplasmic (25S, 18S, 5.8S, and 5S rRNA), Chloroplast (23S, 16S, 5S, and 4.5S), and mitochondrial rRNA (18S and 5S).

**Does the Ribo-Zero Plant kit work on all plant species? What about algae?**\
The Ribo-Zero Plant kit has been tested against Arabidopsis, maize, wheat, rice, corn, and soy bean. In silico testing has also demonstrated compatibility with Japanese honeysuckle (Lonicera japonica), mirror bush (Coprosma repens), moss (Physcomitrella), Tomato (Solanum lycopersicum), Vietnamese coriander (Persicaria odorata), and water clover (Marsilea vistita). Reagents may be compatible with other species as well.

**Does the Ribo-Zero Plant kit work on plant leaves only, or is it also compatible with seeds and roots?**\
The Ribo-Zero Plant kit contains oligos that will remove cytoplasmic and chloroplast rRNA from leaves, seeds and roots.

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| *For any feedback or questions regarding this article (Illumina Knowledge Article #1153), contact Illumina Technical Support* [*techsupport@illumina.com*](mailto:techsupport@illumina.com?subject=Question%2FFeedback%20Regarding%20Illumina%20Knowledge%20Article%20#000001153%20-%20Library%20Preparation%20\&body=Dear%20Illumina%20Technical%20Support,%0D%0A%0D%0A)*.* |


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