03353575910
Roche
DIG Oligonucleotide 3′-End Labeling Kit, 2nd generation
sufficient for 25 labeling reactions (100 pmol of oligonucleotides per assay; 1 ug of a 30-mer oligonucleotide), storage condition avoid repeated freeze/thaw cycles
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About This Item
UNSPSC Code:
41105500
NACRES:
NA.55
Recommended Products
usage
sufficient for 25 labeling reactions (100 pmol of oligonucleotides per assay; 1 ug of a 30-mer oligonucleotide)
Quality Level
manufacturer/tradename
Roche
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General description
The DIG Oligonucleotide 3′-End Labeling Kit, 2nd generation employs the enzyme terminal transferase. It catalyzes the addition of single digoxigenin-labeled dideoxyuridine triphosphates (ddUTP) to the 3′-OH end of oligonucleotides. Thus, helps in labeling oligonucleotides.
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.
Application
DIG Oligonucleotide 3′-End Labeling Kit, 2nd generation is for 3′-end labeling of oligonucleotides from 14 to 100 nucleotides in length with DIG-11-ddUTP and recombinant terminal transferase.
DIG-labeled oligonucleotides has been used in a variety of hybridization techniques:
DIG-labeled oligonucleotides has been used in a variety of hybridization techniques:
- dot/slot blots
- colony/ plaque hybridizations
- Southern blots/ northern blots
- in situ hybridizations
Features and Benefits
- Fast hybridization kinetics, due to the small size of oligonucleotides
- Single-stranded probes, no renaturation during hybridization
- Sequence can be designed according to the experiment
- Specially suited for in situ hybridization; due to their small size, the oligonucleotides readily diffuse into fixed tissues and cells
Packaging
1 kit containing 9 components
Quality
Function tested in a dot blot.
Principle
One DIG-ddUTP molecule is added to the 3′-end of oligonucleotides by recombinant Terminal Transferase. This guarantees a very specific and distinct hybridization signal, which is detected by an enzyme-linked immunoassay with anti-DIG-AP antibody conjugate, and a color or chemiluminescence reaction.
Preparation Note
Activator: sodium sulfate, Tris
Working concentration: Oligonucleotides: 100 pmol
Up to 100 pmol (1 μg of a 30-mer) oligonucleotide can be labeled in a single standard labeling reaction.
Working concentration: Oligonucleotides: 100 pmol
Up to 100 pmol (1 μg of a 30-mer) oligonucleotide can be labeled in a single standard labeling reaction.
Assay Time: The complete procedure from labeling the oligonucleotide to hybridization and detection of the first visible signal can be accomplished within less than 24 hours.
Sample Materials
Oligonucleotides of a length from 14 to 100 nucleotides, purified by HPLC or gel electrophoresis
Sample Materials
Oligonucleotides of a length from 14 to 100 nucleotides, purified by HPLC or gel electrophoresis
Storage and Stability
Store at -15–-25 °C. (unopened kit)
Other Notes
For life science research only. Not for use in diagnostic procedures.
Kit Components Only
Product No.
Description
- Reaction Buffer 5x concentrated
- CoCl<sub>2</sub> Solution 25 mM
- DIG-ddUTP Solution 1 mM
- Recombinant Terminal Transferase 400 U/μl
- Control Oligonucleotide, unlabeled 20 pmol/μl
- Oligonucleotide, DIG-ddUTP labeled 2.5 pmol/μl
- Control DNA, 2.5 pmol/μl pUC 18 DNA, supercoiled
- Glycogen Solution 20 mg/ml
- DNA Dilution Buffer, 50 μg/ml fish sperm DNA
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Danger
Hazard Classifications
Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Aquatic Chronic 2 - Carc. 1B Inhalation - Repr. 1B
Storage Class
6.1D - Non-combustible, acute toxic Cat.3 / toxic hazardous materials or hazardous materials causing chronic effects
wgk_germany
WGK 3
flash_point_f
does not flash
flash_point_c
does not flash
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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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