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LIG1

Sigma-Aldrich

DNA Ligation Kit

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About This Item

Code UNSPSC :
12352204
Nomenclature NACRES :
NA.53

Qualité

for molecular biology

Utilisation

 kit sufficient for 150 ligation reactions

Conditions d'expédition

wet ice

Température de stockage

−20°C

Description générale

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

Composants

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)

Principe

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.

Produit(s) apparenté(s)

Réf. du produit
Description
Tarif

D9380

DNA Polymerase I from Escherichia coli lysogenic for NM 964, buffered aqueous glycerol solution

R0884

T7 RNA Polymerase, recombinant, expressed in E. coli, buffered aqueous solution

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

P4390

Polynucleotide Kinase from T4-infected Escherichia coli, 10 units/μL, buffered aqueous glycerol solution

D7291

Deoxyribonuclease I RNase-free solution from bovine pancreas

R6513

Ribonucléase A from bovine pancreas, for molecular biology, ≥70 Kunitz units/mg protein, lyophilized

R4642

Ribonucléase A from bovine pancreas, (Solution of 50% glycerol, 10mM Tris-HCL pH 8.0)

P6911

Protéase from Streptomyces griseus, BioReagent, DNase, RNase, and nickase, none detected (No RNase.)

LIG2

QuickLink™ DNA Ligation Kit, joins blunt-end and sticky-end DNA fragments

T9285

Tampon Tris-EDTA, 100X concentrate for Northern and Southern blotting

Pictogrammes

Exclamation mark
GHS07

Mention d'avertissement

Warning

Mentions de danger

H319

Classification des risques

Eye Irrit. 2

Code de la classe de stockage

10 - Combustible liquids

Classe de danger pour l'eau (WGK)

WGK 3

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable

Certificats d'analyse (COA)

Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".

Déjà en possession de ce produit ?

Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.

Consulter la Bibliothèque de documents

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)
Lei Wei et al.
Nature microbiology, 5(5), 715-726 (2020-03-11)
Chronic hepatitis B virus (HBV) infections result in 887,000 deaths annually. The central challenge in curing HBV is eradication of the stable covalently closed circular DNA (cccDNA) form of the viral genome, which is formed by the repair of lesion-bearing
Quanxin Long et al.
PLoS pathogens, 13(12), e1006784-e1006784 (2017-12-30)
Hepadnavirus covalently closed circular (ccc) DNA is the bona fide viral transcription template, which plays a pivotal role in viral infection and persistence. Upon infection, the non-replicative cccDNA is converted from the incoming and de novo synthesized viral genomic relaxed
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

Protocoles

DNA Ligation Protocol

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

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