LIG1
DNA Ligation Kit
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About This Item
NACRES:
NA.53
UNSPSC Code:
12352204
grade
Molecular Biology
Quality Level
usage
1 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
Biochem/physiol Actions
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.
Other Notes
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)
signalword
Danger
hcodes
Storage Class
10 - Combustible liquids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
Hazard Classifications
Eye Irrit. 2 - Resp. Sens. 1
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Protocols
Cloning process, joining linear DNA into a vector, is crucial for biotechnological experiments, enabling DNA fragment recombinant technology.