Hieff NGS™ MaxUp Human rRNA Depletion Kit (rRNA & ITS/ETS) is designed to remove rRNA and 45S ITS/ETS from human, mouse and rat total RNA using RNase H-based workflow and to retain mRNA and other non-coding RNA. This kit is suitable for both intact and partially degraded RNA samples (e.g., FFPE RNA). Since degraded FFPE samples usually contain a higher proportion of ITS/ETS than fresh tissue samples, this kit adds probes in the Human 45S ITS/ETS region, and ITS removal can significantly increase the proportion of valid data in raw data.

Features

Strong Specificity: Efficiently removes rRNA as well as ITS/ETS regions from human samples, with excellent performance on challenging FFPE samples.

Broad Input Compatibility: Suitable for a wide range of input amounts (100 ng to 1 μg).

High Removal Efficiency: Achieves >95% removal of rRNA, ITS, and ETS from human, mouse, and rat samples.

Consistent Quality: Strict batch-to-batch performance and stability controls ensure reliable results.

Applications

Gene Expression Research

Alternative splicing analysis

Detection and discovery of non coding RNA

dentification of selective polyadenylation sites

Fusion gene detection

Storage

This product should be stored at -25~-15℃ for 12 months.

Figures

1. Broad Compatibility Across Input Amounts and Sample Types

Figure 2. Efficient rRNA Removal Across Multiple Sample Types and Input Amounts

Library preparation was performed using 12257 and RNA Library Preparation Kit (Cat#12308, with strand-specific protocols) on various RNA sample types—including human, mouse, and rat total RNA—across different input amounts (100 ng to 1 μg). The rRNA residual rate remained consistently low and was significantly reduced compared to Supplier V* across all sample types and input levels.

2. Effective rRNA Removal

Figure 3. Comparison of rRNA Removal Efficiency by Sequencing and qPCR

Using 1 μg of 293T total RNA, samples were processed by oligo-dT enrichment or rRNA depletion. Sequencing measured residual rRNA%(A). qPCR targeting cytoplasmic (18S, 28S, 5.8S) and mitochondrial (12S, 16S) rRNAs showed significant Ct value increases after depletion(B), demonstrating effective rRNA removal.

3. Stability

 Figure 4. Stability Test

(A) 1  μg RNA standard was used to compare rRNA removal efficiency between Yeasen (Cat#12257ES) and Supplier V* after storage at 4 °C for 3 days or freshly prepared. Yeasen’s kit maintained normal removal efficiency and outperformed the competitor. rRNA levels were assessed by qPCR targeting 18S, 28S, and ACT.

(B) RNA standard (547 ng/μL) was fragmented (94 °C, 7 min) and RNA Libraries were prepared using Yeasen Kit (Cat#12308, with strand-specific protocols). rRNA removal remained stable after 11 days at 4 °C, comparable to freshly prepared reagents.

Documents:

Safety Data Sheet

12257_MSDS_HB250711_EN.PDF

Manuals

12257_Manual_HB20260401.pdf

Related Blogs:

LncRNA-Seq Library Preparation Protocol for Human/Mouse/Rat

rRNA Depletion Kit – Efficient rRNA Removal Across All Sample Types