5-(3-Aminoallyl)-dUTP (50 mM)

When incorporated into DNA, Ambion 5-(3-aminoallyl)-dUTP provides a reactive group for addition of other chemical groups. The modified dUTP is深入閱讀

產品號碼	Quantity
AM8439	50 μL

產品號碼 AM8439

價格 (TWD)

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50 μL

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When incorporated into DNA, Ambion 5-(3-aminoallyl)-dUTP provides a reactive group for addition of other chemical groups. The modified dUTP is readily incorporated by reverse transcriptase. The aminoallyl modification enables downstream reaction with amine-reactive compounds such as N-hydroxysuccinimide (NHS) esters; thus aminoallyl-modified cDNA can be labeled with any amine-reactive group moiety.

Use of this modified nucleotide
Use 5-(3-aminoallyl)-dUTP as recommended in your protocol for reverse transcription. Substitution of 30–80% of the dTTP with 5-(3-aminoallyl)-dUTP may be useful, but the exact proportion will depend on experimental goals.

For Research Use Only. Not for use in diagnostic procedures.

Product LineAmbion

Product TypeNucleotides

Purification MethodHPLC

Quantity50 μL

Shipping ConditionDry Ice

Concentration50 mM

For Use With (Application)Nucleic Acid Labeling

FormLiquid

Label or DyeAminoallyl

Unit SizeEach

內容物與存放

5-(3-Aminoallyl)-dUTP, 50 mM (50 μL)

Store at –70°C.

常見問答集 (常見問題)

Why are there only two amino-modified nucleotides in the dNTP mix for cDNA indirect labeling (aminoallyl-dUTP and aminohexyl-dATP) intead of all four nucleotides being modified? Would more modified nucleotides have an adverse effect on the microarray?

Using four modified nucleotides rather than two can cause fluorescence quenching since the fluorophores are positioned very closely to one another. Use of two amino-modified nucleotides in the cDNA synthesis reaction (rather than one) results in a greater incorporation of fluorescent dye and higher signal intensity with small amounts of RNA starting material. Unbiased incorporation of amino-modified dNTPs and the high efficiency of the coupling reaction result in an even distribution of fluorescent signal and high overall levels of fluorescence, increasing the sensitivity and reproducibility of array hybridizations. In addition to quenching, a very high dye density on the cDNA will interfere with hybridization and, therefore, yield lower microarray fluorescence signals. We found that the use of 2 amino-modified nucleotides (at the appropriate concentration) results in optimal incorporation and high microarray signals.

Can dU-containing DNA produced by amplification with dUTP be cut by restriction enzymes?

You will need to confirm with the manufacturer of the restriction enzyme you are using to find out whether or not it can recognize dUTP-containing DNA.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.