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Conjugation Handle Oligonucleotide Modifications

Reactive functional groups for precise attachment of fluorophores, peptides, proteins, antibodies, GalNAc, lipids, polymers, nanoparticles, surfaces, and other payloads.

Click chemistry Amino Thiol Carbonyl reactive Carboxyl Multifunctional Bioorthogonal Custom design

Overview

Conjugation handle modifications introduce reactive functional groups into an oligonucleotide to create a controlled attachment point for downstream bioconjugation. These handles allow DNA, RNA, siRNA, ASOs, aptamers, PNA, and other oligonucleotide formats to be linked to fluorophores, peptides, proteins, antibodies, GalNAc, lipids, polymers, nanoparticles, surfaces, and therapeutic delivery systems.

Bio-Synthesis offers a broad portfolio of functional groups for conjugation, including click chemistry modifiers, amino modifiers, thiol modifiers, carbonyl-reactive groups, carboxyl modifiers, multifunctional handles, bioorthogonal handles, and custom functional group designs. Handles may be incorporated at the 5' end, 3' end, or internal positions depending on the design and chemistry.

Design the handle around the payload

Select the functional group based on the reactive chemistry of the dye, peptide, protein, ligand, surface, polymer, or particle you want to attach.

Conjugation handle oligonucleotide modifications including click chemistry, amino, thiol, carboxyl, carbonyl, and bioorthogonal functional groups

Functional groups for oligonucleotide conjugation, labeling, immobilization, bioorthogonal ligation, and custom bioconjugation design.

Handle • Linker • Payload • QC

Choose the reactive handle first, then tune position, spacer length, purification, and analytical QC to support reliable downstream conjugation.

FG

Functional Group

Click, amino, thiol, carbonyl, carboxyl, multifunctional, and bioorthogonal handle families.

5′

Modification Position

5′, 3′, or internal placement for terminal conjugation, spatial control, or surface display.

PEG

Linker & Spacer

Spacer length can improve accessibility, reduce steric effects, and support conjugation efficiency.

QC

Purification & QC

Project-appropriate purification and analytical review for modified or conjugation-ready oligos.

Select handle

Choose position

Match payload chemistry

Define purification

Confirm QC needs

DNA RNA siRNA ASO Aptamer PNA Peptide conjugates Antibody-oligo conjugates GalNAc PEG / polymer Nanoparticles

Functional Group Selector

Click a conjugation handle category to review typical functional groups, common reactions, and applications. This is the main decision point for choosing an oligo modification for downstream conjugation.

start Here Select the chemistry your payload or workflow needs

7

Handle families

5′ • Internal • 3′

Position options

DNA • RNA • PNA • ASO

Oligo formats

Scientist-Assisted

Design support
N3

Click Chemistry Modifiers

Best for: selective bioorthogonal conjugation when the oligo must be linked to dyes, peptides, antibodies, GalNAc, polymers, nanoparticles, or other payloads after synthesis.

CuAAC SPAAC IEDDA
Recommended handle family

Typical handles

  • Azide
  • Alkyne
  • DBCO
  • BCN
  • Tetrazine
  • TCO

Common uses

  • Fluorophore conjugation
  • Peptide and protein conjugation
  • Antibody-oligo conjugates
  • Targeting ligand attachment

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NH2

Amino Modifiers

Best for: NHS ester coupling, activated ester chemistry, isothiocyanate labeling, and broad attachment to dyes, biotin, surfaces, proteins, and particles.

NHS ester Activated ester Amide coupling
Recommended handle family

Typical handles

  • Amino C6
  • Amino C12
  • Amino serinol
  • Internal amino
  • PEG amino

Common uses

  • Dye labeling
  • Biotin attachment
  • Surface immobilization
  • Protein or enzyme coupling

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SH

Thiol Modifiers

Best for: maleimide coupling, disulfide chemistry, gold surface attachment, nanoparticle functionalization, and thiol-reactive conjugation workflows.

Maleimide Gold binding Disulfide
Recommended handle family

Typical handles

  • Thiol C3
  • Thiol C6
  • Protected thiol
  • Internal thiol

Common uses

  • Gold nanoparticles
  • Biosensors and surfaces
  • PEGylation
  • Protein conjugation

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C=O

Carbonyl-Reactive Functional Groups

Best for: oxime ligation, hydrazone formation, and site-specific conjugation designs involving aldehyde, ketone, hydrazide, or aminooxy chemistry.

Oxime Hydrazone Site-specific
Recommended handle family

Typical handles

  • Aldehyde
  • Ketone
  • Hydrazide
  • Aminooxy

Common uses

  • Site-selective ligation
  • Glycan-related conjugation
  • Special linker design
  • Controlled payload attachment

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2X

Multifunctional Modifiers

Best for: oligos requiring more than one function, such as a targeting ligand plus dye, a capture group plus reactive handle, or orthogonal handles for stepwise conjugation.

Dual function Orthogonal handles Branched designs
Recommended handle family

Typical designs

  • Amino + biotin
  • Azide + fluorophore
  • Azide + thiol
  • Dual click handles
  • Branching handles

Common uses

  • Multifunctional probes
  • Therapeutic research
  • DNA nanotechnology
  • Advanced imaging assays

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COOH

Carboxyl Modifiers

Best for: EDC chemistry, NHS activation, amide bond formation, surface coupling, and attachment to amine-containing partners.

EDC NHS activation Amide bond
Recommended handle family

Typical handles

  • Carboxy C10
  • PEG carboxyl
  • Internal carboxyl
  • Custom carboxyl linkers

Common uses

  • Surface immobilization
  • Protein or peptide attachment
  • Particle coupling
  • Custom linker chemistry

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TCO

Bioorthogonal Handles

Best for: highly selective ligation strategies that minimize interference with biomolecules, complex samples, or sensitive payloads. Useful when fast, orthogonal, or copper-free conjugation is preferred.

Tetrazine ligation Copper-free click Orthogonal pairing
Recommended handle family

Typical handles

  • Tetrazine
  • TCO
  • Norbornene
  • DBCO
  • BCN
  • Cyclooctyne derivatives

Common uses

  • Fast ligand conjugation
  • Antibody-oligo conjugates
  • Cell-compatible labeling strategies
  • Orthogonal multi-step designs

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Need a Custom Functional Group or Linker?

Bio-Synthesis can develop custom linkers, PEG spacers, multifunctional oligonucleotide designs, customer-supplied phosphoramidites, and novel conjugation chemistries for specialized research, diagnostic, and therapeutic applications.

Conjugation Chemistry Quick Guide

Use this quick guide to match the reactive chemistry of your payload or conjugation partner with the appropriate oligonucleotide functional group. This reference helps identify the recommended conjugation handle for common bioconjugation workflows.

If your reagent or workflow uses... Recommended oligo handle Typical result
NHS ester or activated ester Amino modifier Amide-linked dye, biotin, protein, or surface conjugate
Maleimide Thiol modifier Thioether-linked peptide, protein, PEG, or surface conjugate
Azide-bearing payload DBCO, BCN, or alkyne handle Copper-free or copper-assisted click conjugate
Alkyne-bearing payload Azide handle CuAAC click conjugate
Tetrazine ligation TCO or tetrazine handle Fast bioorthogonal ligation pair
EDC coupling Carboxyl or amino handle Amide bond formation
Oxime or hydrazone chemistry Aminooxy, hydrazide, aldehyde, or ketone handle Carbonyl-selective conjugation

Why Choose Bio-Synthesis

Extensive Handle Portfolio

Broad support for standard and specialized reactive functional groups for conjugation-ready oligonucleotides.

Flexible Positioning

Functional groups can be designed for 5', 3', or internal placement depending on chemistry and application.

Custom Chemistry

Support for custom linkers, spacers, multifunctional handles, and customer-supplied building blocks.

Scientist Support

Assistance with handle selection, linker strategy, purification, scale, and downstream conjugation planning.

Frequently Asked Questions

FAQ

What are conjugation handle oligonucleotide modifications?
They are reactive functional groups added to an oligonucleotide to create a defined attachment point for downstream conjugation to dyes, peptides, proteins, antibodies, lipids, polymers, surfaces, or other payloads.
Which functional group should I choose for NHS ester chemistry?
An amino-modified oligo is typically selected for NHS ester or activated ester chemistry because the primary amine forms an amide linkage with activated carboxyl reagents.
When should I choose a thiol-modified oligo?
Thiol-modified oligos are commonly used for maleimide chemistry, gold nanoparticle attachment, biosensor surfaces, disulfide strategies, and thiol-reactive conjugation partners.
Can one oligo contain more than one functional group?
Yes. Dual and multifunctional designs may include combinations such as a dye plus reactive handle, biotin plus amino group, or orthogonal click handles for stepwise conjugation.
Can click chemistry handles be added to DNA or RNA?
Yes. Click-ready handles such as azide, alkyne, DBCO, BCN, tetrazine, and TCO may be considered for DNA, RNA, siRNA, ASO, aptamer, or other oligonucleotide formats depending on design feasibility.
What information helps Bio-Synthesis prepare a quote?
Please provide the oligo sequence, desired functional group, modification position, scale, purification requirement, payload or conjugation partner, intended chemistry, and any linker or spacer preference.

Need help choosing the right oligonucleotide conjugation handle?

Send your oligonucleotide sequence, desired functional group, modification position, payload or conjugation partner, scale, purification requirement, and intended chemistry. Bio-Synthesis can help select the appropriate handle, linker strategy, synthesis format, purification, QC, and downstream conjugation approach.

What to Send

  • Oligonucleotide sequence and format
  • Desired functional group and position
  • Payload or conjugation partner
  • Scale and purification requirement
  • Application and timeline

What We Review

Functional group compatibility, 5′ / 3′ / internal placement, linker spacing, conjugation chemistry, purification strategy, QC needs, and custom handle feasibility.

Why Choose Bio-Synthesis

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