1、Background
rRNA accounts for a high proportion of Total RNA in organisms, but it can provide little effective information. These Rnas will produce a large number of invalid data, which is of little significance for the study of obtaining biological information. Meanwhile, rRNA with a high proportion makes mRNA relatively abundant in transcripts low, thus affecting the detection of transcripts with low abundance. Therefore, in conventional RNA sequencing (RNA-SEQ), the effective removal of rRNA is essential to achieve cost-effective sequencing of RNA.
2、Product description
(1)MaxUp series,rRNA Depletion Kit
The MaxUp series of rRNA removal reagents is based on RNase H digestion to remove ribosomal RNA (rRNA) from a sample's Total RNA to retain messenger RNA (mRNA) and other non-coding Rnas. The sample types are rich, the starting amount is wide, the whole process takes less than 2h, and the manual operation time is less than 10min. At the same time, rRNA can be efficiently removed for both conventional samples and low-quality samples (such as FFPE), significantly improving the effective data information in sequencing data, and realizing cost-effective sequencing of RNA samples.
(2)MaxUp series rRNA removal processes
At present, RNase elimination is the main method of rRNA removal, especially for some RNA degradation samples (such as FFPE samples), RNase elimination can show its advantages.
Figure 1. Schematic diagram of MaxUp series rRNA removal principle
3、Product Performance Display
(1)Plant samples
RNase H was used to digest and remove ribosomal RNA from total RNA from plants, and qPCR was used to compare the expression changes of rRNA gene and mRNA gene before and after removal. The results showed that this product could effectively remove rRNA from plants.
FIG. 2. 1μg Arabidopsis leaf RNA was used for rRNA removal, and qPCR was used to compare the expression changes of rRNA gene and mRNA gene before and after removal
(2)Human/rat/mouse sample
RNase H was used to digest and remove ribosomal RNA from the total RNA from plants, build a library and send sequencing. Data analysis showed that this product could efficiently remove rRNA from such samples.
Figure 3. 1μg total RNA of human, mouse and rat was taken as samples, and the rRNA residues were analyzed by sequencing after rRNA removal
(3)Low quality RNA samples (e.g. FFPE RNA)
Low-quality samples tend to have a large proportion of ITS/ETS sequences (refer to FFPE RNA: DV200=20% in the following figure), and this part of the sequences will also produce a large number of invalid data, so by adding ITS/ETS probes in low-quality samples, the target sequences can be further effectively enriched. In addition to the probe for the conventional 45S Pre-rRNA, the probe design for the Human 45S ITS/ETS region is also added in this product, which ensures that rRNA removal in low-quality samples can be more efficient without affecting the rRNA removal effect in conventional samples.
Figure 4. Comparison of rRNA residues and ITS/ETS residues before and after the addition of ITS/ETS probes
4、Ordering information
Type |
Product name |
Product Code |
Product specifications |
rRNA removal kit |
Hieff NGSTM MaxUp Human rRNA Depletion Kit(rRNA & ITS/ETS) |
12257ES08/24/96 |
8/24/96T |
Hieff NGSTM MaxUp rRNA Depletion Kit (Plant) |
12254ES08/24/96 |
8/24T/96T |