Fluorescence Labeling of Oligonucleotides

As a leading service provider in the field of bioconjugation and drug discovery, BOC Sciences is fully competent and dedicated to serving one-stop oligonucleotide labeling services which has be used in various detection and assay system. BOC Sciences provides a variety of solutions for the preparation of fluorescently labeled oligonucleotides, including labeling regents, independent dyes and custom labeling services.

What is Fluorescently Labeled Oligonucleotides?

Fluorescence labeling of oligonucleotides is the binding of a fluorescent dye to an oligonucleotide molecule for visualization and detection of the oligonucleotide. Fluorescent labeling can be performed by different methods, including direct labeling and indirect labeling. The fluorescent group can be attached to the 5' end, 3' end, base end of nucleoside or phosphate and structure of ribose by covalent bonding according to different needs. After labeling, the fluorescently labeled product needs to be purified for further use. Labeling allows the oligonucleotide to be directly visible under a fluorescence microscope or quantitatively detected by a device such as a fluorometer.

Fig 1. Fluorescently labeled oligonucleotides. (Saneyoshi et al., 2020)

Custom Oligonucleotide Labeling Services

Fluorescent labeled oligonucleotide synthesis

We can prepare your desired fluorescently labeled oligonucleotides by providing the sequence and the type of fluorescent dye required.

Fluorescent labeled oligonucleotide probe design

These probes can be used to detect specific DNA or RNA sequences. You provide the target sequence information and we will design the appropriate probe and fluorescently label it.

Fluorescent labeled oligonucleotide detection

BOC Sciences provides fluorescently labeled oligonucleotide detection services, including fluorescence in situ hybridization and fluorescence quantitative PCR.

Fluorescent labeled oligonucleotide purification

We can provide purification services to purify fluorescently labeled oligonucleotides from mixtures.

Steps for Fluorescent Labeled Oligonucleotides

Step 1 - Synthesize the oligonucleotide

The desired oligonucleotide sequence needs to be synthesized first, the exact sequence length and base composition will depend on experimental needs.

Step 2 - Fluorescent dye labeling

Combine the oligonucleotide with fluorescent dye labeling. Common fluorescent dyes include fluorescein, pyridine dyes, cyanine dyes, etc.

Step 3 - Removal of unreacted fluorescent dyes and oligonucleotides

The reaction needs to be purified to remove unreacted fluorescent dyes and oligonucleotides. Purification can be performed using gel electrophoresis, high performance liquid chromatography, and other methods.

Step 4 - Identification of labeled oligonucleotides

Detect fluorescently labeled oligonucleotides using instruments such as spectrophotometers and fluorescence spectrometers.

Advantages of Fluorescent Labeled Oligonucleotides

• High sensitivity
• High resolution
• Real-time monitoring
• Easy to handle
• Versatility

Application of Fluorescent Labeled Oligonucleotides

Nucleic acid sequencing

At present, most of the gene chips are labeled with fluorescent dyes and scanned by laser confocal chip scanner and CCD chip scanner to determine the results.

Detection of pathogenic pathogens

Known bacteria, viruses, protozoa, parasites and chlamydia, mycoplasma and other microbial nucleic acid sequences, we can design primers and probes to detect by PCR or hybridization to help the diagnosis of diseases.

Diagnosis of genetic disease

With the development of gene mutation detection technology, nucleic acid hybridization, PCR and sequence analysis have gradually become routine methods for the diagnosis of genetic diseases.

Gene expression analysis

Fluorescence quantitative PCR based on fluorescence energy transfer technology can accurately and sensitively determine the concentration of template and detect gene variation, which has become a new method for quantitative detection of nucleic acid.

Our Advantages

• Excellent, high quality, competitive prices
• More than 200 labels to choose from
• Click chemistry allowing completely new dye combinations not available before and so far unreachable labeling densities
• Creating complex labeled hybrid oligonucleotides with the flexibility to be adapted
• Data analysis, detailed report with results and discussion
• All samples are carefully monitored for stability and characterized to ensure batch to batch consistency.

Project Workflow

Frequently Asked Questions (FAQ)

How can I optimize the fluorescent labeling of oligonucleotides for my specific experiment?

Optimization depends on several factors, including the oligonucleotide sequence, the type of fluorophore, and the experimental conditions. We can provide guidance on adjusting the labeling protocol, such as optimizing reaction conditions and selecting the appropriate fluorophore for your detection method.

What should I do if the fluorescent signal of my labeled oligonucleotides is weak or inconsistent?

Weak signals can be caused by improper labeling efficiency or suboptimal fluorophore selection. We recommend checking labeling efficiency through HPLC or adjusting reaction conditions.

How do I avoid nonspecific binding when using fluorescently labeled oligonucleotides?

To minimize nonspecific binding, it's important to optimize reaction conditions, such as salt concentrations and incubation times. We can assist with adjusting these parameters, as well as selecting blocking agents or optimizing the oligonucleotide design for more specific binding.

Can I use fluorescently labeled oligonucleotides in assays with a high concentration of salts or other additives?

Fluorescently labeled oligonucleotides can be used in high-salt conditions, but compatibility depends on the fluorophore used. We can help identify the best fluorophore for your specific assay conditions to ensure optimal performance, even in complex environments.