Recommended Products
grade
for molecular biology
usage
kit sufficient for 150 ligation reactions
shipped in
wet ice
storage temp.
−20°C
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.
Application
Suitable for:
- Joining fragments of DNA into a cloning vector
- Mutagenesis
- Gene anyalysis and structure-function relationships
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)
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.
Signal Word
Warning
Hazard Statements
Precautionary Statements
Hazard Classifications
Eye Irrit. 2
Storage Class Code
10 - Combustible liquids
WGK
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Sambrook, J., et al
Molecular Cloning: A Laboratory Manual, 2, 1-1 (1989)
Ausubel, F. M.
Current Protocols in Molecular Biology, 1, 3-3 (1994)
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Quanxin Long et al.
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Bradley J Eckelmann et al.
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