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- 2025年10月08日
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20
- 保质期：
1年
- 英文名：
EnGen® Spy dCas9 (SNAP-tag®)
- 库存：
999
- 供应商：
上海淳麦
- 规格：
1000U/10000U
EnGen® Spy dCas9 is an inactive mutant of Cas9 nuclease that retains programmable DNA binding activity (1,2). EnGen Spy dCas9 contains two point mutations, D10A and H840A, which inactivate its RuvC and HNH nuclease domains, respectively. The N-terminal SNAP-tag® allows for covalent attachment of fluorophores, biotin, and a number of other conjugates useful for visualization and target enrichment. Like with Cas9 nuclease, DNA binding by EnGen Spy dCas9 requires a guide RNA complementary to a target sequence with a protospacer adjacent motif (PAM), NGG, downstream of the target sequence. EnGen Spy dCas9 contains a SV40 T antigen nuclear localization sequence (NLS) at the C- terminus of the protein.
Product Source
EnGen® Spy dCas9 is expressed as a N-terminal 6XHis-tagged fusion in E. coli.
Reagents Supplied
The following reagents are supplied with this product
| Store at (°C) | Concentration | |
| NEBuffer™ 3.1 | -20 | 10X |
Description
EnGen® Spy dCas9 is an inactive mutant of Cas9 nuclease that retains programmable DNA binding activity (1,2). EnGen Spy dCas9 contains two point mutations, D10A and H840A, which inactivate its RuvC and HNH nuclease domains, respectively. The N-terminal SNAP-tag® allows for covalent attachment of fluorophores, biotin, and a number of other conjugates useful for visualization and target enrichment. Like with Cas9 nuclease, DNA binding by EnGen Spy dCas9 requires a guide RNA complementary to a target sequence with a protospacer adjacent motif (PAM), NGG, downstream of the target sequence. EnGen Spy dCas9 contains a SV40 T antigen nuclear localization sequence (NLS) at the C- terminus of the protein.
Product Source
EnGen® Spy dCas9 is expressed as a N-terminal 6XHis-tagged fusion in E. coli.
Reagents Supplied
The following reagents are supplied with this product
| Store at (°C) | Concentration | |
| NEBuffer™ 3.1 | -20 | 10X |
Advantages and Features
Applications
- Localization of DNA targets in vivo
- Target Enrichment
Properties and Usage
Reaction Conditions
1X NEBuffer 3.1
Incubate at 37°C
1X NEBuffer™ 3.1:
100 mM NaCl
50 mM Tris-HCl
10 mM MgCl2
100 μg/ml BSA
pH 7.9 @ 25°C
Storage Temperature
-20°C
Storage Conditions
300 mM NaCl
10 mM Tris-HCl
0.1 mM EDTA
1 mM DTT
50% Glycerol
pH 7.4 @ 25°C
Related Products
Companion Products
- EnGen® sgRNA Synthesis Kit, S. pyogenes
- HiScribe™ T7 Quick High Yield RNA Synthesis Kit
- SNAP-Vista® Green
- SNAP-Capture Magnetic Beads
Notes
- 1..20 µM is equal to ~3.6 mg/ml and 1uM is equal to ~ 0.18 mg/ml EnGen® Spy dCas9.
- 2. This product is compatible with Diluent Buffer B: 300 mM NaCl, 10 mM Tris-HCl, 0.1 mM EDTA, 1 mM DTT, 500 µg/ml BSA and 50% glycerol. (pH 7.4 @ 25°C).
References
- Qi, LS., et. al. (2013). Cell. 152 (5), 1173-83.
- Qi, LS., et. al. (2013). Cell. 154 (2), 442-51.
上海淳麦生物科技有限公司专注于感受态细胞、质粒、载体、农杆菌、酵母等生命科学领域的产品研发和技术服务支持。
上海淳麦生物科技有限公司在感受态细胞的研发方面，实力雄厚，拥有了克隆、表达、酵母和农杆菌4大类，共计百余种感受态细胞。在电击感受态方面，拥有了大肠杆菌和农杆菌电击感受态种类，极大提高了客户在转基因以及文库构建方面的成功率。
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文献和实验DNA extraction from Mutation Detection Enhancement (MDE) Gel Stained with Silver Nitrate
extension cycle at 72 ˚C for 5 min. PCR reactions were performed in Gene Amp® PCR System 480 (Perkin-Elmer, Norwalk, CT, USA). DNA samples of normal subjects (blood donors) were included as well as negative control (with no DNA sample) in each reaction.
The ribonuclease protection assay (RPA)
, 5X transcription buffer, and RPA template set to RT. For each probe synthesis, add the following in order to a 1.5 ml Eppendorf tube: 1 µl RNasin® 1 µl GACU pool 2 µl DTT 4 µl 5X transcription buffer 1 µl RPA Template Set 10 µl [a-32
ORNL MICROARRAY HYBRIDIZATION PROTOCOLS
Direct labeling of total RNA with Cy3 and Cy5: A. MATERIALS RNeasy® Mini Kit (Qiagen; Cat # 74106) SuperScript II RT (200U/µL) (Life Technologies; Cat # 18064-014) QIAquick PCR Purification Kit (Qiagen; Cat # 28106)
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