Aminoallyl-dUTP - Solution

For general laboratory use.

Envio: shipped on gel packs

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

Validade: 12 months after date of delivery

Fórmula molecular: C12H20N3O14P3 (free acid)

Peso molecular: 523.22 g/mol (free acid)

CAS#: 116840-18-7

Pureza: ≥ 95 % (HPLC)

Forma: solution in water

Concentração: 100 mM - 110 mM

pH: 7.5 ±0.5

Propriedades espectroscópicas: λmax 289 nm, ε 7.1 L mmol-1 cm-1 (Tris-HCl pH 7.5)

Formulários:
Incorporation into DNA/cDNA by

• PCR with Taq polymerase^{[1], in-house data}
• Nick Translation with DNAse I/ DNA Polymerase I^[2]
• Primer Extension with Klenow exo^{- [3]}
• 3'-End Labeling with Terminal deoxynucleotidyl Transferase (TdT)^[4]
• Reverse Transcription with MMLV Reverse Transcriptase^[2]

Descrição:
Aminoallyl-dUTP is recommended for two-step labeling of DNA/cDNA e.g. by PCR, Nick Translation, Primer Extension, 3'-End Labeling and Reverse Transcription. It is enzymatically incorporated into DNA/cDNA as substitute for its natural counterpart dTTP. The resulting Amine-functionalized DNA/cDNA can subsequently be labeled via the classic Amine/NHS Ester reaction that offers the choice to introduce a Biotin group (via NHS Ester of Biotin) for subsequent purification tasksto introduce fluorescent group (via NHS Ester of fluorescent dyes) for subsequent microscopic imaging

Referências selecionadas:
[1] Dirsch et al. (2007) Probe production for in situ hybridization by PCR and subsequent covalent labeling with fluorescent dyes. Appl. Immunohistochem. Mol. Morphol. 3:332.

[2] Cox et al. (2004) Fluorescent DNA hybridization probe preparation using amine modification and reactive dye coupling. BioTechniques 36:114.

[3] Cherkasov et al. (2010) New Nucleotide Analogues with Enhanced Signal Properties. Bioconjugate Chem. 21 (1):122.

[4] Unciti-Broceta et al. (2003) The use of solid supports to generate nucleic acid carriers.Accounts of Chemical Research 45:1140.