General description
Digoxigenin-11-dUTP, alkali-stable is provided in 1 mM tetralithium salt, solution.
Application
Digoxigenin-11-dUTP, alkali-stable has been used to label probes for
fluorescent in situ hHybridization (FISH) and polymerase chain reaction (PCR)
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.
Biochem/physiol Actions
Digoxigenin (DIG)-11-deoxyuridine triphosphate (dUTP) replaces deoxythymidine triphosphate (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 polymerase chain reaction (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.
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.
Quality
Typical analysis: At least 85% DIG-11-dUTP (HPLC, area%).
Function tested by RPL.
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.
Other Notes
For life science research only. Not for use in diagnostic procedures.
- UPC:
- 51283510
- Condition:
- New
- Availability:
- 3-5 Days
- Weight:
- 1.00 Ounces
- HazmatClass:
- No
- MPN:
- 11558706910
