DIG-11-dUTP

For general laboratory use.

Shipping: shipped on gel packs

Storage Conditions: store at -20 °C
Short term exposure (up to 1 week cumulative) to ambient temperature possible.

Shelf Life: 12 months after date of delivery

Molecular Formula: C₄₃H₆₅N₄O₂₁P₃ (free acid)

Molecular Weight: 1066.91 g/mol (free acid)

Exact Mass: 1066.34 g/mol (free acid)

Purity: ≥ 95 % (HPLC)

Form: filtered solution (30 kDa) in 10 mM Tris-HCl

Color: colorless to slightly yellow

Concentration: 1.0 mM - 1.1 mM

pH: 7.5 ±0.5

Spectroscopic Properties: λ_max 290 nm, ε 8.8 L mmol^-1 cm^-1 (Tris-HCl pH 7.5)

Applications:
Incorporation into DNA/cDNA by
- PCR with Taq polymerase ^{[1,2] & in-house data}
- Nick Translation with DNAse I/ DNA Polymerase I ^{[3] and in-house data}
- Primer Extension with Klenow exo^{- [4]}
- 3’-End Labeling with Terminal deoxynucleotidyl Transferase (TdT) ^[5]
- Reverse Transcription with MMLV Reverse Transcriptase ^[6]
Incorporation into RNA by
- 3’-End Labeling with Terminal deoxynucleotidyl Transferase (TdT) ^[7]

Description:
Digoxigenin-11-dUTP is enzymatically incorporated into DNA/cDNA as substitute for its natural counterpart dTTP. The resulting Digoxigenin-labeled DNA/cDNA probes are subsequently detected using Digoxigenin-antibodies conjugated with horseradish peroxidase (HRP), alkaline phosphatase (AP) or a fluorescent dye. Optimal substrate properties and thus labeling efficiency is ensured by a 11-atom linker attached to the C5 position of uridine.

Recommended Digoxigenin-11-dUTP/dTTP ratio for PCR and Nick Translation: 35% Digoxigenin-11-dUTP/ 65% dTTP

Please note: The optimal final concentration of Digoxigenin-11-dUTP may very depending on the application and assay conditions. For optimal product yields and high incorporation rates an individual optimization of the Digoxigenin-11-dUTP/dTTP ratio is recommended.

Related products:

• HighFidelity Digoxigenin PCR Labeling Kit, #APP-101-DIGX
• Digoxigenin NT Labeling Kit, #PP-310-DIGX

BIOZ Product Citations:

Powered by Bioz See more details on Bioz

Selected References:
[1] Anderson et al. (2005) Incorporation of reporter-labeled nucleotides by DNA polymerases. Biotechniques 38:257.
[2] Jackson et. al (1991) Detection of Shiga Toxin-Producing Shigella dysenteriae Type 1 and Escherichia coli by Using Polymerase Chain Reaction with Incorporation of Digoxigenin-11-dUTP. J Clin Microbiol. 29 (9):1910.
[3] Dauwerse et al. (1999) Two-colour FISH detection of the inv (16) in interphase nuclei of patients with acute myeloid leukemia. Br J Haematol 106:111.
[4] Wiegant et al. (2008) Probe Labeling and Fluorescence In Situ Hybridization. Current Protocols in Cytometry: Unit 8.3.
[5] Schmitz et al. (2008) Nonradioactive labeling of oligonucleotides in vitro with the hapten digoxigenin by tailing with terminal transferase. Anal Biochem 199:222.
[6] Grimmond et al. (2001) Expression Profiling with cDNA Microarray's: A User's Perspective and Guide. In: DNA Microarrays: Gene Expression Applications (Jordan). Springer Verlag Berlin Heidelberg.
[7] Rosemeyer et al. (1995) Nonradioactive 3'-end-labeling of RNA molecules of different lengths by terminal deoxynucleotidyltransferase Anal Biochem 224:446.