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LIG1 Sigma

DNA Ligation Kit

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Properties

Related Categories Cell Biology, Circuit Design and Metabolic Pathway Engineering, Modifying Enzymes, Molecular Biology, Molecular Biology Enzymes,
grade   for molecular biology
usage   1 kit sufficient for 150 ligation reactions
shipped in   wet ice
storage temp.   −20°C

Description

Components

Sufficient for 150 reactions:
• 300uL 10X Ligation Buffer (D2176) in 250 mM Tris-HCl (pH 7.8) 100 mM MgCl2, and 10 mM dithiothreitol
• 3 x 100 units T4 DNA Ligase (D2886) in 50% glycerol with 10 mM Tris-HCl (pH 7.5) 50 mM KCl, and 1 mM dithiothreitol
• 3 x 100 uL 10 mM ATP ( A3702)
• 50 uL Control pBR322 DNA, HAE III Digest (D9430) 0.5 ug/ul in 10 mM Tris-HCl (pH 8.0), and 1 mM EDTA
• 1.5 mL 24% (w/v) PEG Solution, (P 2454)
• 1.5 mL Molecular Biology Grade Water (W4502)

General description

Sigma’s DNA Ligation Kit contains all of the reagents necessary to perform DNA ligation reactions. Sticky-end ligations are more efficient than blunt-end ligations; these can be facilitated with the addition of PEG.

Principle

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.

Application

Suitable for:
• Joining fragments of DNA into a cloning vector
• Mutagenesis
• Gene anyalysis and structure-function relationships

Price and Availability

Safety & Documentation

Safety Information

Symbol 
GHS07  GHS07
Signal word 
Warning
Hazard statements 
Precautionary statements 
RIDADR 
NONH for all modes of transport
Protocols & Articles

Protocols

DNA Ligation Kit

Product Description Reagents Provided Precautions and Disclaimer Storage Procedure Materials References
Keywords: AGE, Cloning, Electrophoresis, Enzyme activity, Gel electrophoresis, Gene expression, Molecular biology, Nucleic acid annealing, Peptide synthesis, transformation

Peer-Reviewed Papers
15

References

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D8276 DNA Polymerase I, Klenow Fragment from Escherichia coli, buffered aqueous glycerol solution
D2886 DNA Ligase from T4-infected Escherichia coli, buffered aqueous glycerol solution
P4978 Phosphatase, Alkaline from calf intestine, buffered aqueous glycerol solution
T4427 Terminal Transferase from calf thymus, buffered aqueous glycerol solution
P4390 Polynucleotide Kinase from T4-infected Escherichia coli, 10 units/μL, buffered aqueous glycerol solution
E1131 Exonuclease III from Escherichia coli BE25 /psGR3, buffered aqueous glycerol solution
D7291 Deoxyribonuclease I RNase-free solution from bovine pancreas
T7428 Topoisomerase I from wheat germ, buffered aqueous glycerol solution
R6513 Ribonuclease A from bovine pancreas, for molecular biology, ≥70 Kunitz units/mg protein, lyophilized
R4642 Ribonuclease A from bovine pancreas, (Solution of 50% glycerol, 10mM Tris-HCL pH 8.0)

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