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Nick translation DNA labeling system

Nick translation system for preparing FISH probes
 
ENZ-42910 50 Reactions 278.00 USD
Bulk Request
 
The Nick Translation DNA Labeling System provides a simple and efficient method for generating labeled DNA. The complete system includes all of the necessary reagents required for 50 Nick translation reactions and is recommended for labeling of double stranded DNA larger than 1kb for fluorescent in situ hybridization (FISH) applications. The Nick Translation DNA Labeling System can accommodate a wide range of biotin-labeled and fluorophore-labeled nucleotides. In addition to choice of label, the kit design allows the user to optimize incorporation and product size by adjusting the ratio of labeled-dUTP to dTTP and DNase I concentration respectively.
ENZ-42910 specific activity
Column purified labeled FISH probes demonstrate consistent incorporation and specific activity. The nick translation reaction enables rapid and consistent production of labeled DNA, accommodates a wide range of labeled nucleotides and is ideal for multiplexed applications.
ENZ-42910 yield
Enzo Life Sciences' Nick Translation DNA Labeling System allows for consistent production of labeled DNA probes. Column purified probes produced using BAC DNA (1 ug) demonstrate significant yields and show no bias among labeled nucleotides. Any variance between the amount of starting material and labeled probe is attributed to loss during column purification.
ENZ-42910 tubes
SYSTEM OVERVIEW. The Enzo Life Sciences' Nick Translation DNA Labeling System contains individualized components, including DNase I, DNA Polymerase I and labeled dUTP. This allows optimization beyond the recommended reaction protocol included with the kit. DNase I introduces randomly distributed nicks to double stranded DNA at low enzyme concentrations. The 5' → 3' exonuclease activity of Polymerase I removes nucleotides from the 3' side of the nick while synthesizing a partial new complementary strand using the 3'-OH termini as primer. In the presence of dye- or biotin-labeled dUTP, Polymerase I incorporates labeled dUTP instead of dTTP. The well-balanced nuclease/polymerase activities of the enzyme mix ensure the generation of highly labeled double stranded DNA fragments.
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ENZ-42910 specific activity ENZ-42910 yield ENZ-42910 tubes

Product Specification

Quality Control:Performance Testing: A sample kit from each lot of Nick Translation DNA Labeling System is used to label 1.0µg BAC DNA with Orange 552 dUTP (Prod. No. ENZ-42842) and hybridized to metaphase spreads using the protocols described in the user manual.
Minimum Specifications:
- Electrophoresis of the reaction on a non-denaturing agrose gel must yield a distinct smear between 50 to 500 base pairs.
- An average of 100 picomoles of labeled nucleotide must be incorporated per μg of sample DNA as described by the method in the user manual.
- A fluorescent labeled locus-specific FISH probe generated from 1 µg BAC DNA must demonstrate chromosome specific hybridization confirmed by a labeled CEN probe.
 
Use/Stability:Stable for at least one year after receipt when stored as recommended.
 
Handling:Avoid freeze/thaw cycles.
 
Long Term Storage:-20°C
 
Kit/Set Contains:100 µL, DNase I 180 µL, dTTP 250 µL, DNA Polymerase I 250 µL, dNTP Mix 250 µL, Reaction Buffer 250 µL, Stop Buffer 1 mL, DNase I Dilution Buffer 10 mL, Nuclease-free Water
 

Product Literature References

Imprinted expression of UBE3A in non-neuronal cells from a Prader-Willi syndrome patient with an atypical deletion: K. Martins-Taylor, et al.; Hum. Mol. Genet. 23, 2364 (2014), Application(s): Labeling DNA for FISH, Abstract; Full Text
Altered expression of MGMT in high-grade gliomas results from the combined effect of epigenetic and genetic aberrations: J. Ramalho-Carvalho, et al.; PLoS One 8, e58206 (2013), Application(s): Labeling DNA for FISH, Abstract; Full Text
Common fusion transcripts identified in colorectal cancer cell lines by high-throughput RNA sequencing: T. Nome, et al.; Transl. Oncol. 6, 546 (2013), Application(s): Labeling DNA for FISH, Abstract; Full Text

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