上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。
产品信息
产品来源
大肠杆菌菌株,携带有克隆自化脓性链球菌(Streptococcus pyogenes)的 Cas9 基因,在其编码蛋白的 N 端和 C 端都含有猴病毒 40(SV40)核定位序列(NLS),N 端同时带有 6xHis 标签。
- 产品类别:
- Genome Editing Products,
- Programmable Nucleases Products
- 应用:
- Genome Editing Applications
-
产品组分信息
本产品提供以下试剂或组分:
NEB # 名称 组分货号 储存温度 数量 浓度 -
M0646T -20 EnGen® Spy Cas9 NLS M0646TVIAL -20 1 x 500 pmol 20 µM NEBuffer™ r3.1 B6003SVIAL -20 1 x 1.25 ml 10 X
-
M0646M -20 EnGen® Spy Cas9 NLS M0646MVIAL -20 1 x 2,500 pmol 20 µM NEBuffer™ r3.1 B6003SVIAL -20 1 x 1.25 ml 10 X
-
-
特性和用法
反应条件
1X NEBuffer™ r3.1
Incubate at 37°C1X NEBuffer™ r3.1
100 mM NaCl
50 mM Tris-HCl
10 mM MgCl2
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)贮存溶液
300 mM NaCl
10 mM Tris-HCl
0.1 mM EDTA
1 mM DTT
50% Glycerol
pH 7.4 @ 25°C热失活
65°C for 5 minutes
-
注意事项
- 20 μM 等于 3.22 mg/ml
-
参考文献
- Jinek M. et al. (2012). Science. 816-821. DOI: 10.1126/Science.1225829. Epub 2012 Jun 28 PubMedID: 22745249,
操作说明、说明书 & 用法
-
操作说明
- In vitro digestion of DNA with EnGen Spy Cas9 NLS (M0646)
- Protocol for Electroporation of EnGen® SpyCas9 NLS, S. pyogenes RNP (ribonucleoprotein) into adherent cells using the Lonza 4D-Nucleofector™ System
- Transfection of Cas9 RNP (ribonucleoprotein) into adherent cells using the Lipofectamine® RNAiMAX
- Electroporation of EnGen® Spy Cas9 NLS RNP (ribonucleoprotein) into adherent cells using the Neon® Electroporation System
- In vitro digestion of plasmid DNA with EnGen SpRY Cas9 (NEB #M0669)
FAQs & 问题解决指南
-
FAQs
- Where is the nuclear localization signal on EnGen® Spy Cas9 NLS located?
- Which nuclear localization signal is fused to EnGen® Spy Cas9 NLS?
- What is the difference between EnGen® Spy Cas9 NLS (NEB #M0646) and Cas9 NLS, S.pyogenes (NEB #M0641)?
- Why do I observe incomplete digestion?
- Why does digestion/editing efficiency differ between two different guide RNAs?
- Does NEB provide plasmids for gRNA cloning?
- How do I dilute the enzyme to 1 μM for in vitro reactions?
- Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?