一般描述
Sigma′s DNA Ligation Kit contains all of the reagents necessary to perform DNA ligation reactions at room temperature using blunt or sticky ends. This kit replaces previous workflows requiring cooking steps and long incubations. Pre-made buffers allow for fast & easy setup.
应用
QuickLink™ DNA Ligation Kit has been used for the ligation of pGL2-basic luciferase-reporter plasmid with vascular endothelial growth factor (VEGF) promoter sequence and for the ligation of NLR family caspase activating and recruitment domain (CARD)-containing protein 4 (Nlrc4) promoter with the luciferase plasmid vector pGL4.10-luc2
The QuickLink™ DNA Ligation Kit is suitable for:
- Joining blunt or cohesive-end fragments of DNA into a cloning vector
- Recircularization of linear DNA
- Formation of concatamers
- dsDNA nick repair
特点和优势
- Fast 5 minutes ligation
- High ligation efficiency
- Room temperature reactions – no cooling required
- Perform bacterial transformation with the reaction mixture
组分
Sufficient for 50 reactions:
- 500uL 2X Ligation Buffer A (L9537)
- 100uL 5X Ligation Buffer B (L9662)
- 250 units T4 DNA Ligase (D2886) in 50% glycerol with 10 mM Tris-HCl (pH 7.5) 50 mM KCl, and 1 mM DTT
原理
One of the most important steps in the cloning process is the ligation of linear DNA into a cloning vector. DNA ligations are performed by incubating DNA fragments with appropriately linearized cloning vectors in the presence of buffer, ATP, and DNA ligase. Many parameters affect ligations such as the relative ratio of insert to vector, the quality and type of the DNA ends, the temperature of ligation and the concentration of DNA.
法律信息
QuickLink is a trademark of Sigma-Aldrich Co. LLC
相关产品
产品编号
说明
价格
储存分类代码
10 - Combustible liquids
闪点(°F)
Not applicable
闪点(°C)
Not applicable
法规信息
新产品
K Hayashi et al.
Nucleic acids research, 14(19), 7617-7631 (1986-10-10)
Polyethylene glycol (PEG) stimulates ligation with T4 DNA ligase. In 10% (w/v) PEG 6,000 solutions, only intermolecular ligation is enhanced by monovalent cations, while both inter- and intramolecular ligation occur without their presence. Similar stimulation was also caused by divalent
ErbB2 overexpression in mammary cells upregulates VEGF through the core promoter
Loureiro RMB, et al.
Biochemical and Biophysical Research Communications, 326(2), 455-465 (2005)
R Rossi et al.
Nucleic acids research, 25(11), 2106-2113 (1997-06-01)
ATP-dependent DNA ligases are essential enzymes in both DNA replication and DNA repair processes. Here we report a functional characterization of the T4 DNA ligase. One N-terminal and two C-terminal deletion mutants were expressed in Escherichia coli as histidine- tagged
I R Lehman
Science (New York, N.Y.), 186(4166), 790-797 (1974-11-29)
DNA ligase of E. coli is a polypeptide of molecular weight 75,000. The comparable T4-induced enzyme is somewhat smaller (63,000 to 68,000). Both enzymes catalyze the synthesis of phosphodiester bonds between adjacent 5'-phosphoryl and 3'-hydroxyl groups in nicked duplex DNA
Constitutive Activation of the Nlrc4 inflammasome prevents hepatic fibrosis and promotes hepatic regeneration after partial hepatectomy
DeSantis DA, et al.
Mediators of Inflammation, 2015(2), 455-465 (2015)
实验方案
T4 DNA ligase is used for the joining of DNA molecules with compatible cohesive (sticky) termini, joining of blunt ended double stranded DNA molecules
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