grade
Molecular Biology
Quality Level
usage
1 kit sufficient for 150 ligation reactions
shipped in
wet ice
storage temp.
−20°C
General description
Sigma 的 DNA 连接试剂盒包含进行 DNA 连接反应所需的所有试剂。粘性末端连接比平末端连接更有效。加入 PEG 可促进这些反应。
Application
适用于:
- 将 DNA 片段连接到克隆载体中
- 诱变
- 基因分析与结构功能关系
Biochem/physiol Actions
克隆过程中最重要的步骤之一是将线性 DNA 连接到克隆载体中。在缓冲液、ATP 和 DNA 连接酶的存在下,通过将 DNA 片段与适当线性化的克隆载体孵育可进行 DNA 连接。插入物与载体的相对比例、DNA 末端的质量和类型、连接温度和 DNA 浓度等多种参数会影响连接。
Other Notes
足以进行 150 次反应:
- 在 250 mM Tris-HCl(pH 7.8),100 mM MgCl2 和 10 mM 二硫苏糖醇中的 300uL 10X 连接缓冲液(D2176)
- 含有 3 x 100 单位 T4 DNA 连接酶(D2886),10 mM Tris-HCl,pH 7.5,50 mM KCl 和 1 mM 二硫苏糖醇的 50% 甘油溶液
- 3 x 100 uL 10 mM ATP(A3702)
- 50μL 对照 pBR322 DNA,HAE III Digest(D9430)浓度为 0.5 ug/ul,溶于 10 mM Tris-HCl(pH 8.0)和 1 mM EDTA 中
- 1.5 mL 24%(w/v)PEG 溶液,(P 2454)
- 1.5 mL 分子生物学级水(W4502)
存储类别
10 - Combustible liquids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
signalword
Danger
hcodes
Hazard Classifications
Eye Irrit. 2 - Resp. Sens. 1
法规信息
常规特殊物品
此项目有
实验方案
Cloning process, joining linear DNA into a vector, is crucial for biotechnological experiments, enabling DNA fragment recombinant technology.
Sambrook, J., et al
Molecular Cloning: A Laboratory Manual, 2, 1-1 (1989)
Lei Wei et al.
Nature microbiology, 5(5), 715-726 (2020-03-11)
Chronic hepatitis B virus (HBV) infections result in 887,000 deaths annually. The central challenge in curing HBV is eradication of the stable covalently closed circular DNA (cccDNA) form of the viral genome, which is formed by the repair of lesion-bearing
Quanxin Long et al.
PLoS pathogens, 13(12), e1006784-e1006784 (2017-12-30)
Hepadnavirus covalently closed circular (ccc) DNA is the bona fide viral transcription template, which plays a pivotal role in viral infection and persistence. Upon infection, the non-replicative cccDNA is converted from the incoming and de novo synthesized viral genomic relaxed
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