Education, Technology
The Basic Components of Labeling an Oligo
September 4, 2015 by admin · Leave a Comment
Summary: Here is a brief guide to the main components of the labeling process.
Labeling an oligonucleotide can be achieved through a radioactive approach, using chemiluminescent methods or by fluorescent label techniques. When you look in-depth into the labeling process, there are three main components that you should know: a space, a signalling moiety, and a reactive group.
A spacer is what separates the luminescent moiety from DNA within the molecules and also has the ability to draw a fine line between hydrophilicity and hydrophobicity. What it also does in terms of the label itself is that it can modify the flexibility and spacing of the label in regards to the DNA.
What is a signalling moiety? It’s either a molecule like a fluorophore, fluorescein or an enzyme. Each type caters to a different type of labeling technique. For instance, fluorescein is used via direct labeling and enzymes are used for indirect labeling. It’s important to note which molecule falls with their compatible labeling type.
The reactive group is what attaches the label itself to the oligonuclotide. One of the most important components of the entire process, it generates the means of conjunction.
Some common types of labeling reactions include: the reaction of an alkyne-modified oligonucleotide with an azide-modified label to end up creating a triazole linkage, and also the reaction of an azide-modified oligo with an alkyne-modified label to create, also, an azide-modified label.
When it comes to multiple labels, there is a process that utilizes dual-labeled probes. Also known as the hydrolysis probe, multiple dyes label the oligonucleotide and through hydrolysis, an increase in fluorescence is received.
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