Pular para o conteúdo
Merck
Todas as fotos(1)

Key Documents

11745816910

Roche

DIG-Nick Translation Mix

greener alternative

sufficient for 40 labeling reactions, pkg of 160 μL, suitable for hybridization

Sinônimo(s):

nick translation

Faça loginpara ver os preços organizacionais e de contrato


About This Item

Código UNSPSC:
41105500

forma

solution

Nível de qualidade

uso

sufficient for 40 labeling reactions

embalagem

pkg of 160 μL

fabricante/nome comercial

Roche

características do produto alternativo mais ecológico

Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.

sustainability

Greener Alternative Product

técnica(s)

hybridization: suitable

categoria alternativa mais ecológica

temperatura de armazenamento

−20°C

Descrição geral

DIG-Nick Translation Mix is a convenient enzyme and nucleotide mixture for nick translation. It utilizes a combination of DNase and DNA Polymerase to nick one strand of the DNA helix, then incorporates labeled nucleotides as the polymerase examines, or "proofreads" the nicked site. Large plasmids, cosmids and polymerase chain reaction (PCR) fragments are all regularly used with this method and the DNA can be linearized or supercoiled. Individual templates produce consistent results in the standard 90-minutes reaction and result in an average probe length of 200 bp up to 500 bp. The molar ratio of digoxigenin (DIG)-11-deoxyuridine triphosphate (dUTP) to deoxythymidine triphosphate (dTTP) is adjusted to ensure that every 20th to 25th nucleotide in the newly synthesized DNA is modified with DIG. This density of haptens in the DNA yields the highest sensitivity in the immunological detection reaction.
We are committed to bringing you Greener Alternative Products, which adhere to one or more of The 12 Principles of Greener Chemistry. This product is designed as a safer chemical.  The DIG System was established as a sensitive and cost-effective alternative to radioactivity for the labeling and detection of nucleic acids. There are many available publications that prove the versatility of the DIG System, so use of radio-labeling is no longer the only option for labeling of DNA for hybridization.

Especificidade

Heat inactivation: Stop the reaction by adding 1μl 0.5M EDTA (pH 8.0) and heating to 65°C for 10 minutes.

Aplicação

DIG-Nick Translation Mix has been used for the generation of highly sensitive probes for in situ hybridization labeled with Digoxigenin-11-dUTP.
Note: The mix can also be used for filter hybridization techniques, however, for highly sensitive filter hybridization probes, we recommend that you use DIG-High Prime from Roche Applied Science.
For nonradioactive labeling of in situ probes with other haptens and fluorophores, Roche Applied Science offers the Biotin-Nick Translation Mix and the Nick Translation Mix (without nucleotides).

Qualidade

Function tested in a dot spot assay.

Princípio

The nick translation method is based on the ability of DNase I to introduce randomly distributed nicks into DNA at low enzyme concentrations in the presence of MgCl2.
E. coli DNA Polymerase I synthesizes DNA complementary to the intact strand in a 5′→3′ direction using the 3′-OH termini of the nick as a primer. The 5′→3′ exonucleolytic activity of DNA polymerase I simultaneously removes nucleotides in the direction of synthesis. The polymerase activity sequentially replaces the removed nucleotides with isotope-labeled or hapten-labeled deoxyribonucleoside triphosphates. At low temperature (+15°C), the unlabeled DNA in the reaction is thus replaced by newly synthesized labeled DNA.

Nota de preparo

1 vial with 5x concentrated stabilized reaction buffer in 50% glycerol (v/v) and DNA Polymerase I, DNase I, 0.25mM dATP, 0.25mM dCTP, 0.25mM dGTP, 0.17mM dTTP and 0.08mM DIG-11-dUTP (alkali-stable).
Assay Time: 100 minutes
Sample Materials: Supercoiled and linearized plasmid DNA, Supercoiled and linearized cosmid DNA, Purified PCR products.
Note: Denaturing of the template before nick translation is not required.

Outras notas

For life science research only. Not for use in diagnostic procedures.

Código de classe de armazenamento

12 - Non Combustible Liquids

Classe de risco de água (WGK)

WGK 1

Ponto de fulgor (°F)

No data available

Ponto de fulgor (°C)

No data available


Certificados de análise (COA)

Busque Certificados de análise (COA) digitando o Número do Lote do produto. Os números de lote e remessa podem ser encontrados no rótulo de um produto após a palavra “Lot” ou “Batch”.

Já possui este produto?

Encontre a documentação dos produtos que você adquiriu recentemente na biblioteca de documentos.

Visite a Biblioteca de Documentos

Haifeng Cui et al.
Planta, 229(2), 323-330 (2008-11-06)
Regenerates were obtained following somatic hybridization between tall wheatgrass (Agropyron elongatum) and bread wheat (Triticum aestivum cv. Jinan177) protoplasts. Two lines (CU and XI) were self-fertile in the first (R0) and subsequent (R1 and R2) generations. The phenotype of each
Liang Meng et al.
PloS one, 9(10), e107922-e107922 (2014-10-02)
Testis-specific protein kinase 1 (Tesk1) is a serine/threonine kinase with unique structural features. In the present study, we cloned and characterized the tesk1 gene of tongue sole, Cynoglossus semilaevis. The full-length tesk1 cDNA consists of 1,672 nucleotides, encoding a 331
Allopolyploid speciation of the Mexican tetraploid potato species Solanum stoloniferum and S. hjertingii revealed by genomic in situ hybridization.
Pendinen G, et al.
Genome / National Research Council Canada = Genome / Conseil National de Recherches Canada, 51, 714-720 (2008)
Towards a reliable method to select potentially high value Erianthus hybrids.
Piperidis N, et al.
International Sugar Journal, 1-9 (2013)
Massimiliano Rampin et al.
Comparative cytogenetics, 6(3), 287-300 (2012-01-01)
Genomic In Situ Hybridization (GISH) is a powerful tool to identify and to quantify genomic constituents in allopolyploids, and is mainly based on hybridization of highly and moderate repetitive sequences. In animals, as opposed to plants, GISH has not been

Artigos

Digoxigenin (DIG) labeling methods and kits for DNA and RNA DIG probes, random primed DNA labeling, nick translation labeling, 5’ and 3’ oligonucleotide end-labeling.

Nossa equipe de cientistas tem experiência em todas as áreas de pesquisa, incluindo Life Sciences, ciência de materiais, síntese química, cromatografia, química analítica e muitas outras.

Entre em contato com a assistência técnica