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BioReagent,Endonuclease,EnzymoPure™,for DNA and RNA applications,PCR试剂,分子生物学级,无DNA酶,无支原体,核酸外切酶,用于DNA合成,用于体外转录,通过内毒素测试,重组 级 提供 ≥95%(SDS-PAGE) 纯度,适用于对基线干扰要求严格的色谱和分析工作流程。
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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
RNase R(Ribonuclease R)是一种来源于大肠杆菌 RNR 超家族且依赖 Mg 2+的核糖核酸外切酶,从3'→5'方向将线性RNA完全水解成二核苷酸和三核苷酸。对环状 RNA 及双链 RNA 不敏感,可用于特殊结构 RNA 的生产,如环状RNA(circRNA)、套索结构 RNA(lariat RNA)、3'突出末端少于 7 个核苷酸的双链 RNA 以及具有复杂结构的 tRNA 等。RNase R常用于基因表达和可变剪切研究,可消化线性 RNA 使 circ RNA 或 lariat RNA 得到富集。
| R749973 | Component | 500U | 2KU | 10KU | Storage |
| R749973A | RNase R (20U/μL) | 25 μL | 100 μL | 500 μL | -20°C. Avoid freeze/thaw cycle |
| R749973B | 10×RNase R Buffer | 500 μL | 2×1 mL | 10×1 mL | -20°C. Avoid freeze/thaw cycle |
| R749973C | RNase R Dilution Buffer | 500 μL | 2×1 mL | 10×1 mL | -20°C. Avoid freeze/thaw cycle |
来源
重组 E.coli
活性单位定义
在 37℃标准反应条件下, 10min 将 1μg poly(A)转化为酸溶性核苷酸所需的酶量定义为1U
储存缓冲液: 50mMTris-HCl (pH7.5 at 25℃), 100mM NaCl, 0.1mM EDTA, 1mM DTT, 0.1%Triton X-100 (v/v), 50%Glycerol (v/v).
反应缓冲液: 200 mM Tris-HCl (pH 8.0 at 25°C), 1 M KCl, 1 mM MgCl₂.
推荐 RNase R 消化体系:
|
反应条件:37℃消化 15min,70℃孵育 10min 可使酶失活。
*配制反应体系时,RNase R(20U/μL)可用RNase R Dilution Buffer稀释成合适的工作浓度,建议现配现用。
适用范围
去除线性 RNA,从生物样本中富集 circ RNA,可变剪切研究,内含子套索序列的分析和鉴定等。
注意事项
1.RNase R 活性的发挥需要 0.1-1.0mM Mg 2+。
2.随底物 RNA 的增加,可适当延长消化时间和增加酶量。
3.RNA 样本中 EDTA 含量可能会影响 RNase R 的活性。 试减少 RNase R 用量或缩短消化时间。
4.有些 circRNA 或套索结构 RNA 在经长时间 RNase R 消化会导致丰度降低,可能是因为其耐受RNase R 消化能力弱,可尝试减少 RNase R 用量或缩短消化时间 。
Product presentation:
RNase R (Ribonuclease R) is an MG-2 + dependent exonuclease derived from the Escherichia coli RNR superfamily that completely hydrolyzes linear RNA into dinucleotides and trinucleotides from the 3'→5' direction. It is not sensitive to circular RNA and double-stranded RNA, and can be used for the production of special structural RNA, such as circRNA, lariat RNA, double-stranded RNA with 3' protruding end less than 7 nucleotides, and tRNA with complex structure. RNase R is commonly used in gene expression and variable shear studies, and can digest linear RNA to enrich circ RNA or lariat RNA.
| R749973 | Component | 500U | 2KU | 10KU | Storage |
| R749973A | RNase R (20U/μL) | 25 μL | 100 μL | 500 μL | -20°C. Avoid freeze/thaw cycle |
| R749973B | 10×RNase R Buffer | 500 μL | 2×1 mL | 10×1 mL | -20°C. Avoid freeze/thaw cycle |
| R749973C | RNase R Dilution Buffer | 500 μL | 2×1 mL | 10×1 mL | -20°C. Avoid freeze/thaw cycle |
Source
Recombinant E.coli
Activity unit definition
The amount of enzyme required to convert 1μg poly(A) to acid-soluble nucleotides for 10min under standard reaction conditions at 37 ° C is defined as 1U.
Recommended RNase R digestive system:
Storage buffer: 50mMTris-HCl (pH7.5 at 25℃), 100mM NaCl, 0.1mM EDTA, 1mM DTT, 0.1%Triton X-100 (v/v), 50%Glycerol (v/v).
Reaction Buffer: 200 mM Tris-HCl (pH 8.0 at 25°C), 1 M KCl, 1 mM MgCl₂.
Recommended RNase R digestive system:
|
Reaction conditions: digestion at 37℃ for 15min, incubation at 70℃ for 10min can deactivate the enzyme.
* When preparing the reaction system, RNase R (20U/μL) can be diluted to a suitable working concentration by RNase R Dilution Buffer, and it is recommended to use it now.
Scope of application
Removal of linear RNA, enrichment of circ RNA from biological samples, variable shear studies, analysis and identification of intron lasso sequences, etc.
Matters needing attention
1. The play of RNase R activity requires 0.1-1.0mM Mg 2+.
2. With the increase of substrate RNA, the digestion time and enzyme amount can be appropriately prolonged.
3. The content of EDTA in RNA samples may affect the activity of RNase R. Try to reduce the amount of RNase R or shorten the digestion time.
4. The abundance of some circRNA or lasso structure RNA will decrease after prolonged digestion of RNase R, which may be due to their weak digestion ability to tolerate RNase R. Therefore, attempts can be made to reduce the amount of RNase R or shorten the digestion time.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | R749973 | |
| 分析证书 | R749973 | |
| 分析证书 | R749973 | |
| 分析证书 | R749973 |
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