the DIG High Prime DNA labeling and Detection Initiation Kit I uses the steroid hapten digoxin (DIG) to label DNA probes by enzyme-immunoassay for hybridization and subsequent color rendering. The kit includes ready-to-use sealer, a comprehensive reserve consisting of nitrotetrazolium Blue (NBT) / 5-bromo-4-chloro-3-indolyl phosphate (BCIP and DIG Easy Hyb particles). The DIG-High Prime mixture consists of stable Klenow enzymes, nucleotides, primers and reaction buffers - all dissolved in a convenient reagent. The sample material can be: at least 100bp DNA fragments, linearized plasmids, cosmid or λDNA, or superhelical DNA "random-bound" DNA labeling methods first developed by Feinberg and Vogelstein are based on hybridizing oligonucleotides of all possible sequences into denatured DNA to be labeled. The initial addition of DNA serves only as a template for the synthesis of labeled DNA and does not degrade during the reaction, thus allowing very small amounts of DNA (10ng) to be labeled. Complementary DNA strands were synthesized by Klenow polymerase using the 3 '-OH terminal of random oligonucleotides as primer. Modified deoxyribonucleoside triphosphate with digitoxigenin labeling present in the reactants is incorporated into newly synthesized complementary DNA strands. This convenient kit enables the detection of DIG-labeled hybridization products with color development by immediately triggering labeled DNA templates from alkali-unstable DIG-11-dUTP. We are committed to providing you with more environmentally friendly alternatives that meet one or more of the requirements of the 12 Principles of Green Chemistry. This product is a safe chemical. The DIG system is a sensitive and cost-effective alternative to nucleic acid radiolabelling and detection. There are many published papers demonstrating the versatility of the DIG system, so the use of radiolabels is no longer the only option for labeling DNA for hybridization.

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DIG-High Prime DNA marker and detection initiation Kit I has been used in a variety of hybridization techniques:

Southern blot

Nrthern mark

Spot imprinting

Colony and plaque hybridization

All types of filter hybridization

Single-copy genetic testing of total genomic DNA can even detect samples of highly complex organisms, such as humans, barley and wheat

The yield of DIG-labeled probes was high after incubation for one hour or overnight.

Characteristics and advantages

We are committed to providing you with more environmentally friendly alternatives that meet one or more of the requirements of the 12 Principles of Green Chemistry. This product is a safe chemical. The DIG system is a sensitive and cost-effective alternative to radiolabelling and radiological methods for nucleic acid detection. Many existing publications demonstrate the versatility of the DIG system, so the use of radiolabelling is no longer the only option for hybridization DNA marking.

package

One kit contains 7 components.

Quality

Function has been tested by dot hybridization test.

Other instructions

For life science research only. Not used for diagnostic operations.

Kit component only

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 GranμLes