Digoxigenin-11-dUTP, alkali-stable

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

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.

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.

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.

Typical analysis: At least 85% DIG-11-dUTP (HPLC, area%).
Function tested by RPL.

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.

For life science research only. Not for use in diagnostic procedures.