Application
Digoxigenin-11-dUTP, alkali-stable, is suitable for nonradioactive DNA labeling (e.g., random priming, PCR labeling, tailing, or nick translation). DIG-11-dUTP replaces dTTP in the random-primed DNA labeling reaction or in nick translation in a ratio of 35% DIG-11-dUTP and 65% dTTP. DIG-11-dUTP can also replace dTTP in labeling during PCR (e.g., with Taq DNA Polymerase), 3′-tailing (with Terminal Transferase) and cDNA synthesis (e.g., with Transcriptor or M-MuLV Reverse Transcriptase). It is ideal as a substrate for DNA polymerase,Taq DNA polymerase, Terminal Transferase and reverse transcriptase.
- Labeled DNA can be subsequently detected with the:DIG Nucleic Acid Detection Kit
- DIG Luminescent Detection Kit
Note: When this nucleotide is subjected to 0.1 - 0.5 NaOH at +15 to +25°C, the DIG moiety will remain attached. It is therefore ideal for use in DIG-labeled DNA that needs to survive exposure to alkaline treatment. However, if you intend to make a DIG-labeled probe that can be removed by alkali denaturation (e.g ., during stripping and reprobing of blots), do not use this alkali-stable preparation. Instead, use the alkali-labile form of DIG-11-dUTP.
Features and Benefits
Digoxigenin is bound to deoxyuridine triphosphate via an alkali-resistant ether linkage.
Resistance to alkali: The preparation is stable in the presence of 0.1 - 0.5M NaOH at +15 to +25°C.
General Description
Digoxigenin-11-dUTP is alkali-stable and comprises a 1 mM solution of the tetralithium salt.
Other Notes
For life science research only. Not for use in diagnostic procedures.
Preparation Note
Working solution: We recommend to prepare a deoxynucleotide mix containing dATP, dCTP, dGTP, dTTP (10 mM, each); (e.g., for the preparation of 100 μl nucleotide mix add 10 μl of dATP, dCTP, dGTP, dTTP (each) to 60 μl Water, PCR Grade).
Storage conditions (working solution): A decomposition of approx. 5% may occur within 4 weeks at 15 to 25 °C.
Quality
Typical analysis: At least 85% DIG-11-dUTP (HPLC, area%). Function tested by RPL.
Assay: ≥85% (HPLC)
Form: solution
Mol. Wt.: Mr 1090.7
Mfr. No.: Roche
Packaging: pkg of 125 μL (11558706910 [1 mM]); pkg of 25 μL (11093088910 [1 mM]); pkg of 5 × 125 μL (11570013910 [5x1 mM])
Absorption.: εmax 22430 and 8780; A 222 and 292 nm
Storage Temp.: −20°C