Biotin Labeled Oligonucleotides

BOC Sciences' custom biomolecule labeled oligonucleotide services include experimental design as well as the development of protocols and procedures for preparing the conjugates requested by our customers. As a leading service provider in the field of drug discovery and research, we have accumulated extensive experience with biotinylated oligonucleotides, and can offer our customers a wide portfolio of products.

Fig.1 Oligonucleotide biotinylation reactions. (Vito, 2020)

What are Biotin Labeled Oligonucleotides?

Biotin-labeled oligonucleotides are compounds in which a biotin molecule is attached to an oligonucleotide molecule. Biotin is a member of the B complex of vitamins and has a high affinity and specificity for interacting with other molecules. In biological research, biotin-labeled oligonucleotides are commonly used to detect and isolate specific nucleic acid sequences or proteins associated with nucleic acids.

The biotin part can be connected to oligonucleotides by 8-atomic spacer (Biotin-on), 15-atomic spacer (Biotin-TEG) or thymidine residue (Biotin-DT). Biotin can be connected to the 5' end, the 3' end, or can be used internally in the sequence. Biotin molecules linked to the 3' end can also be used to prevent 3' extension in the 3' exonuclease digestion and amplification reaction. The biotin used for 3cm-labeling is C3 biotin or 3cm-biotin-TEG.

The Advantages of Biotin Labeled Oligonucleotides

• The corresponding sequences can be synthesized according to the need, the chain is short, the complexity is low, and the hybridization time is short.
• The change of one base in the target sequence can be identified, because the base mismatch in the short probe can greatly reduce the Tm value of the hybrid.
• It can be synthesized in large quantities, making the probe cheap and non-radioactive labeling by enzymatic or chemical methods.

Biotin Labeled Oligonucleotides Services

BOC Sciences provides biotin-labeled oligonucleotide services, including the design of oligonucleotide sequences, preparation, purification, and validation of biotin-labeled oligonucleotides.

Oligonucleotide sequence design

The target nucleic acid sequence is determined according to the research needs. Sequence specificity and complementarity need to be considered during the design process to ensure that the target sequence is accurately recognized and bound in the sample.

Preparation of biotin-labeled oligonucleotides

• Synthetic biotin-labeled oligonucleotides are usually synthesized using chemical synthesis techniques.
• Selection of one end of the oligonucleotide to introduce an amino modification is accomplished by dissolving the appropriate modifying reagent in a solvent containing alkaline conditions and reacting it with the oligonucleotide.
• The selected biotin labeling reagent is subjected to an activation process, usually using NHS (N-hydroxysuccinimide) as the activation reagent. NHS-biotin or biotin-PEG-NHS is dissolved in an organic solvent.
• The activated biotin labeling reagent is reacted with an amino-modified oligonucleotide. The activated reagent solution is added to the amino-modified oligonucleotide solution and the reaction is carried out under appropriate pH and temperature conditions. The reaction time can be optimized for the specific reagent and oligonucleotide.

Labeling validation and purification

The completion of the reaction is confirmed by nucleic acid analysis techniques and biotin binding affinity assays. In addition, purification of the labeled oligonucleotides is required to remove possible impurities.

Optimization and validation applications

Application optimization and validation of biotin-labeled oligonucleotides are performed to ensure optimal signal strength and specificity.

Applications of Biotin Labeled Oligonucleotides

Biotin-modified oligonucleotides bind tightly to streptavidin proteins, which can be labeled with fluorescent dyes and enzymes or as intermediate conjugates for attachment to the biological surface of solids, and different molecular biology and purification methods incorporate biotin modification. Biotin modifications can be added to the 5' or end of the oligonucleotide using either the C6 or TEG (tetra-ethyleneglycol, 15 atom) intermediate arm, the biotin TEG needs to be purified, and the intermediate biotin modification can be added via the dT base, a form that requires more purification steps. The primers are biotin-labeled and can be used in non-radioactive immunoassays to detect proteins, intracellular chemical staining, cell isolation, nucleic acid isolation, hybridization to detect specific DNA/RNA sequences, and ion channel conformational changes.

Our Advantages

• Extensive experience in the fields of biotin, nucleic acid chemistry and dye chemistry
• Obtaining more stable biotin or streptavidin complex
• Advanced instruments and equipment
• Analytical HPLC and MS analyses are performed in every development cycle.
• Guaranteed Quality: All samples are carefully monitored for stability and characterized to ensure batch to batch consistency.
• Full spectrum of Modification and Activation Services

Project Workflow