Description

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 based on 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: specifically remove rRNA and ITS/ETS from human samples, especially for FFPE samples
• High compatibility of template starting amount: applicable to 100 ng~1μg sample
• High removal effect: for rRNA, ITS and ETS in human\mouse and rat samples, the removal effect is more than 95%
• Stable quality: strict batch performance and stability quality control

Applications

• Gene Expression Research
• Alternative splicing analysis
• Detection and discovery of non coding RNA
• dentification of selective polyadenylation sites
• Fusion gene detection

Specifications

Depletion Technology	RNase H
Sample Type	Total RNA of human, mouse and rat
Final Product Type	mRNA and other non coding RNAs
No. of Reactions	24/96 Preps
Starting Material Amount	100 ng~1 μg total RNA
Target	Remove rRNA and 45S ITS/ETS form human total RNA

Components

Components No.	Name	12257ES24 (24T)	12257ES96 (96T)
12257-A	Hybridization Buffer	72 μL	288 μL
12257-B	Probe Mix (rRNA & ITS/ETS)	48 μL	192 μL
12257-C	RNase H Buffer	72 μL	288 μL
12257-D	RNase H	48 μL	192 μL
12257-E	DNase I Buffer	660 μL	2×1320 μL
12257-F	DNase I	60 μL	240 μL

Shipping and Storage

All the components are shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.

Figures

• rRNA removal specificity

For conventional samples, rRNA and ITS/EST can be removed by more than 98%, while for FFPE samples with integrity of about 50%, rRNA and ITS/EST can be removed by more than 95%. At the same time, FFPE with poor integrity can also be removed effectively.

Table 1. rRNA Removal specificity effect

RNA quality	Removal scheme	Input	rRNA (%)	ITS/ETS (%)	Mapping (%)
293T RNA;  RIN=10	remove rRNA	1μg	0.19	4.6	98.28
	remove rRNA and ITS/ETS	1μg	0.15	0.04	98.79
FFPE RNA;  DV200 =90%	remove rRNA	500ng	0.23	4.75	95.35
	remove rRNA and ITS/ETS	500ng	0.21	0.02	95.42
FFPE RNA;  DV200 =50%	remove rRNA	500ng	1.4	16.28	95.57
	remove rRNA and ITS/ETS	500ng	0.56	0.11	95.51
FFPE RNA;  DV200 =20%	remove rRNA	1μg	0.35	67.61	58.4
	remove rRNA and ITS/ETS	1μg	0.79	0.84	60.35

Note: Different human RNA samples were used to remove rRNA or rRNA and ITS/ETS, and to construct libraries with RNA library Prep kit after ITS/ETS. The proportions of rRNA and ITS/ETS in offline data were analyzed by sequencing.

• Sequencing data performance

Table 2. Sequencing data performance

Sample	DV200	Kit	Clean Q20 (%)	Clean Q30 (%)	Clean GC (%)	rRNA (%)	ITS/ETS (%)	unique(%)
1	24%	YEASEN Cat#12257	96.75	92.68	57.05	4.58	2.42	96.90
2	40%		96.56	92.42	55.31	1.93	0.63	96.45
3	53%		97.4	93.52	45.19	0.23	0.01	98.08
4	76%		97.67	93.78	51.28	0.44	0.01	93.63