추천 제품
Grade
for molecular biology
Quality Level
형태
liquid
사용
kit sufficient for 50 ligation reactions
기술
molecular cloning: suitable
배송 상태
dry ice
저장 온도
−70°C
일반 설명
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.
애플리케이션
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
특징 및 장점
- Fast 5 minutes ligation
- High ligation efficiency
- Room temperature reactions – no cooling required
- Perform bacterial transformation with the reaction mixture
성분
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
원리
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.
법적 정보
QuickLink is a trademark of Sigma-Aldrich Co. LLC
관련 제품
제품 번호
설명
가격
Storage Class Code
10 - Combustible liquids
Flash Point (°F)
Not applicable
Flash Point (°C)
Not applicable
시험 성적서(COA)
제품의 로트/배치 번호를 입력하여 시험 성적서(COA)을 검색하십시오. 로트 및 배치 번호는 제품 라벨에 있는 ‘로트’ 또는 ‘배치’라는 용어 뒤에서 찾을 수 있습니다.
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
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)
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
프로토콜
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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