11585606910
Roche
DIG-High Prime
sufficient for 40 labeling reactions, pkg of 160 μL, solution
Synonym(s):
DIG system
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About This Item
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form
solution
Quality Level
usage
sufficient for 40 labeling reactions
packaging
pkg of 160 μL
manufacturer/tradename
Roche
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storage temp.
−20°C
General description
Specificity
Application
Due to highly specific and sensitive detection systems, DIG-labeled DNA probes can be used for single-copy gene detection in 1μg total human DNA. The use of the alkali-labile form of DIG-dUTP, in which the digoxigenin moiety is connected to the spacer arm via an alkali-labile ester bond, enables easier and more efficient stripping and reprobing of blots.
Features and Benefits
- DNA amounts ranging from 10ng to 3μg in a standard reaction.
- DNA of different lengths ranging from small restriction fragments to λ or cosmid DNA.
- DNA, supercoiled or linearized.
- DNA in low melting-point agarose.
Labeling efficiency:
A standard labeling reaction with 1μg template yields 0.8μg newly synthesized digoxigenin-labeled DNA after 1 hour, and 2μg after a 20-hour incubation at +37°C.
Contents
5x concentrated random primer mix: 1U/ μl Klenow polymerase, labeling grade, 1mM dATP, 1mM dCTP, 1mM dGTP, 0.65mM dTTP, 0.35mM DIG-11-dUTP, alkali labile in 50% (v/v) glycerol.
Quality
In a standard assay with 1μg linearized pBR 328, 0.8μg of DIG-labeled DNA is synthesized after 1 hour, and 2.3μg after 20 hours. When this labeled DNA is used for hybridization at a concentration of 20ng/ml, 0.03pg homologous DNA are detected by chemiluminescence with the anti-DIG-alkaline phosphatase conjugate and CSPD on a dot or Southern blot.
Principle
Preparation Note
Assay Time: 80 minutes
Sample Materials
- DNA fragments of at least 100bp
- Linearized plasmid, cosmid or λDNA
- Supercoiled DNA
- Or minimal amounts of DNA (10ng), e.g., DNA restriction fragments isolated from gels or in molten agarose
Note: To obtain optimal results, template DNA should be linearized and should have a size of = 100bp or larger. Template DNA > 5kb should be restriction-digested using a 4bp cutter prior to labeling.
Other Notes
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
No data available
Flash Point(C)
No data available
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Articles
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.
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