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Roche

DIG-High Prime DNA Labeling and Detection Starter Kit I

greener alternative

sufficient for 12 labeling reactions, sufficient for 24 blots, suitable for hybridization, suitable for Northern blotting, suitable for Southern blotting

Synonym(s):

DIG system, dna labeling

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About This Item

UNSPSC Code:
41105500

usage

sufficient for 12 labeling reactions
sufficient for 24 blots

Quality Level

packaging

kit of 1 (7 components)

manufacturer/tradename

Roche

greener alternative product characteristics

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

sustainability

Greener Alternative Product

technique(s)

Northern blotting: suitable
Southern blotting: suitable
hybridization: suitable

greener alternative category

storage temp.

−20°C

General description

The DIG High Prime DNA Labeling and Detection Starter Kit I uses digoxigenin (DIG), a steroid hapten, to label DNA probes for hybridization and subsequent color detection by enzyme immunoassay. This kit contains a ready-made blocking solution, combined stock solution of of nitroblue tetrazolium (NBT)/ 5-bromo-4-chloro-3-indolyl-phosphate (BCIP) and DIG Easy Hyb granules. The DIG-High Prime mixture includes stabilized Klenow enzyme, nucleotides, primers and reaction buffer, all in one convenient reagent. The sample material can be: DNA fragments of at least 100bp, linearized plasmid, cosmid or λDNA, or supercoiled DNA. The "random primed" DNA labeling method originally developed by Feinberg and Vogelstein is based on the hybridization of oligonucleotides of all possible sequences to the denatured DNA to be labeled. The input DNA serves as a template for the synthesis of labeled DNA and is not degraded during the reaction, making it possible to label minimal amounts of DNA (10ng) with this method. In this method, the complementary DNA strand is synthesized by Klenow polymerase using the 3′-OH termini of the random oligonucleotides as primers. Modified deoxyribonucleoside triphosphates, labeled with digoxigenin, present in the reaction are incorporated into the newly synthesized complementary DNA strand. This is a convenient kit for random-primed labeling of DNA templates with DIG-11-dUTP, alkali-labile, and color detection of the DIG-labeled hybrids. 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.

Application

DIG-High Prime DNA Labeling and Detection Starter Kit I has been used in a variety of hybridization techniques:
  • in Southern blots
  • in northern blots
  • in dot blots
  • in colony and plaque hybridizations
  • for all types of filter hybridization
  • for single-copy gene detection in total genomic DNA, even from organisms with high complexity, for example, human, barley and wheat
DIG-labeled probes are generated at high yield within one hour or after overnight incubation.

Features and Benefits

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 using 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.

Packaging

1 kit containing 7 components.

Quality

Function tested in a dot blot.

Other Notes

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

Kit Components Only

Product No.
Description

  • DIG-High Prime 5x concentrated

  • DIG-labeled Control DNA, pBR328 (linearized with Bam HI) 5 μg/ml

  • DNA Dilution Buffer

  • Anti-Digoxigenin-AP Conjugate antibody

  • NBT/BCIP Stock Solution, concentrated

  • Blocking Solution 10x concentrated

  • DIG Easy Hyb Granules

Pictograms

Exclamation mark

Signal Word

Warning

Hazard Statements

Hazard Classifications

Eye Irrit. 2 - Skin Irrit. 2

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 3

Flash Point(F)

does not flash

Flash Point(C)

does not flash


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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