Aminoallyl nucleotide

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  Aminoallyl-UTP structure

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  Heatmap using aminoallyl nucleotide

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  Heck coupling aminoallyl nucleotide reaction

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  Tagging modified uridine

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  Process of preparing amino allyl-labelled cDNA

Aminoallyl Nucleotide

Aminoallyl nucleotides are modified nucleotides that contain an aminoallyl group attached to the nucleotide base. These modified nucleotides are commonly used in molecular biology and biochemistry for labeling nucleic acids with fluorescent dyes or other detectable markers.

Structure

Aminoallyl nucleotides are derived from standard nucleotides by the addition of an aminoallyl group. This group is typically attached to the base of the nucleotide, such as adenine, cytosine, guanine, or uracil. The aminoallyl group provides a reactive amine that can be used for conjugation with various labels.

Applications

Aminoallyl nucleotides are primarily used in the labeling of DNA and RNA for various applications, including:

• Microarray Analysis: Aminoallyl nucleotides are incorporated into cDNA during reverse transcription. The aminoallyl-modified cDNA can then be labeled with fluorescent dyes for use in DNA microarrays.
• Fluorescent In Situ Hybridization (FISH): These nucleotides can be used to label probes for FISH, allowing for the visualization of specific DNA sequences within chromosomes.
• Quantitative PCR (qPCR): Aminoallyl nucleotides can be used to label probes or primers in qPCR assays, facilitating the detection and quantification of target sequences.

Synthesis

The synthesis of aminoallyl nucleotides involves the chemical modification of standard nucleotides. Typically, the aminoallyl group is introduced through a chemical reaction that attaches the group to the nucleotide base. This process requires careful control to ensure that the nucleotide retains its ability to pair with complementary bases.

Advantages

Aminoallyl nucleotides offer several advantages in molecular biology applications:

• Versatility: The reactive amine group allows for the attachment of a wide range of labels, including fluorescent dyes, biotin, and other markers.
• Stability: The modified nucleotides are generally stable and can be incorporated into nucleic acids without disrupting their function.
• Efficiency: The labeling process is efficient, allowing for high levels of incorporation and strong signal generation.

Limitations

While aminoallyl nucleotides are highly useful, they also have some limitations:

• Complex Synthesis: The chemical synthesis of aminoallyl nucleotides can be complex and requires specialized reagents and conditions.
• Potential for Interference: The presence of the aminoallyl group may interfere with some biological processes, although this is generally minimal.

Related Pages

• Nucleotide
• DNA
• RNA
• Fluorescent dye
• Microarray
• Polymerase chain reaction