Not available outside of the UK & Ireland.

Application

Digoxigenin-11-dUTP, alkali-stable has been used to label probes forfluorescent 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.

General description

Digoxigenin-11-dUTP, alkali-stable is provided in 1 mM tetralithium salt, solution.

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.