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MilliporeSigma

LIG1

DNA Ligation Kit

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NACRES:
NA.53
UNSPSC Code:
12352204
Service technique
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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)


pictograms

Health hazard

signalword

Danger

hcodes

Classe de stockage

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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Protocoles

Cloning process, joining linear DNA into a vector, is crucial for biotechnological experiments, enabling DNA fragment recombinant technology.


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)
Bradley J Eckelmann et al.
NAR cancer, 2(3), zcaa013-zcaa013 (2020-08-11)
Homologous recombination/end joining (HR/HEJ)-deficient cancers with BRCA mutations utilize alternative DNA double-strand break repair pathways, particularly alternative non-homologous end joining or microhomology-mediated end joining (alt-EJ/MMEJ) during S and G2 cell cycle phases. Depletion of alt-EJ factors, including XRCC1, PARP1 and



Numéro d'article de commerce international

RéférenceGTIN
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HPA041431-100UL04061836326272
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