Frequently asked questions regarding the Ribo Zero Plant and Globin kits

What molecules are removed with the Ribo-Zero Globin kit? The Ribo-Zero removal mix targets cytoplasmic rRNA, mitochondrial rRNA, plus globin mRNA.

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

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. The only component that differs is the rRNA Removal Mix.

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.

Does the Ribo-Zero Plant kit work on all plant species? What about algae? 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.

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

Do you recommend using the Ribo-Zero Globin kit on any sample 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. Illumina recommends the use of the Ribo-Zero Gold kit for samples other than blood.

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.

Can samples generated with the plant or globin TruSeq protocols be multiplexed in the same lane as human TruSeq RNA samples? Yes, but the indexes are the same. Customers should ensure that all samples have different index combinations.

For any feedback or questions regarding this article (Illumina Knowledge Article #1153), contact Illumina Technical Support [e-mail].