Wszystkie zdjęcia(1)
Kluczowe dokumenty
11745816910
Roche
DIG-Nick Translation Mix
sufficient for 40 labeling reactions, pkg of 160 μL, suitable for hybridization
Synonim(y):
tłumaczenie nicka
Zaloguj sięWyświetlanie cen organizacyjnych i kontraktowych
About This Item
Kod UNSPSC:
41105500
Polecane produkty
Formularz
solution
Poziom jakości
zastosowanie
sufficient for 40 labeling reactions
opakowanie
pkg of 160 μL
producent / nazwa handlowa
Roche
charakterystyka ekologicznej alternatywy
Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.
sustainability
Greener Alternative Product
metody
hybridization: suitable
kategoria ekologicznej alternatywy
, Aligned
temp. przechowywania
−20°C
Opis ogólny
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.
Specyficzność
Heat inactivation: Stop the reaction by adding 1μl 0.5M EDTA (pH 8.0) and heating to 65°C for 10 minutes.
Zastosowanie
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).
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).
Jakość
Function tested in a dot spot assay.
Zasada
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.
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.
Uwaga dotycząca przygotowania
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.
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.
Inne uwagi
For life science research only. Not for use in diagnostic procedures.
Ta strona może zawierać tekst przetłumaczony maszynowo.
Kod klasy składowania
12 - Non Combustible Liquids
Klasa zagrożenia wodnego (WGK)
WGK 1
Temperatura zapłonu (°F)
No data available
Temperatura zapłonu (°C)
No data available
Wybierz jedną z najnowszych wersji:
Masz już ten produkt?
Dokumenty związane z niedawno zakupionymi produktami zostały zamieszczone w Bibliotece dokumentów.
Klienci oglądali również te produkty
Prachi N Ghule et al.
Proceedings of the National Academy of Sciences of the United States of America, 105(44), 16964-16969 (2008-10-30)
Human embryonic stem (hES) cells have an abbreviated G(1) phase of the cell cycle. How cells expedite G(1) events that are required for the initiation of S phase has not been resolved. One key regulatory pathway that controls G(1)/S-phase transition
Production of hexaploid triticale by a synthetic hexaploid wheat-rye hybrid method.
Hao M, et al.
Euphytica, 193, 347-357 (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
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
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
Produkty
Metody znakowania digoksygeniną (DIG) i zestawy do sond DNA i RNA DIG, znakowanie DNA z losowym primerem, znakowanie nickiem translacyjnym, znakowanie końcowe oligonukleotydów 5' i 3'.
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.
Nasz zespół naukowców ma doświadczenie we wszystkich obszarach badań, w tym w naukach przyrodniczych, materiałoznawstwie, syntezie chemicznej, chromatografii, analityce i wielu innych dziedzinach.
Skontaktuj się z zespołem ds. pomocy technicznej