Stable, high-binding PNAs for Diagostic Applications

Request a Quote Today Speak to a Scientist

Rapid. Reliable. Secure

Custom PNA Synthesis

High-affinity, nuclease-resistant PNAs tailored to your assay, diagnostic, or therapeutic research.

Overview

Peptide Nucleic Acids (PNAs) are synthetic DNA analogs where the sugar-phosphate backbone is replaced by a neutral N-(2-aminoethyl)-glycine peptide-like backbone. The neutral charge reduces electrostatic repulsion and enables exceptional sequence-specific binding to DNA and RNA.

Bio-Synthesis offers end-to-end custom PNA synthesis, from sequence design to labeling and conjugation, delivered with rigorous QC documentation. Scales range from milligrams to grams with optional advanced testing.

Speak to a Scientist

At-a-Glance

  • Length: 5–40 mer (longer by request)
  • Purification: HPLC standard; PAGE optional
  • Positions: N-terminus, C-terminus, internal labeling
  • Labels: dyes, biotin, haptens; linkers: PEG, spacers; click handles
  • Conjugations: peptide–PNA, oligo–PNA, antibody–PNA
  • QC: MALDI-TOF MS, analytical HPLC; optional melting/functional assays

Peptide Nucleic Acid (PNA)

Peptide Nucleic Acids (PNAs) are synthetic analogs of DNA in which the traditional sugar–phosphate backbone is replaced with a peptide-like structure. First developed in 1991 by Dr. Peter E. Nielsen, Michael Egholm, Rolf Berg, and Ole Buchardt, PNAs feature an uncharged, achiral backbone composed of repeating N-(2-aminoethyl)-glycine units connected by amide bonds.

The nucleobases adenine (A), cytosine (C), guanine (G), and thymine (T) are attached to this backbone via methylene carbonyl linkages, preserving their ability to form Watson–Crick base pairs. Despite their radically different structure, PNAs hybridize to complementary DNA or RNA sequences with high specificity and affinity.

  • Faster hybridization kinetics
  • Exceptional sequence discrimination (enhanced mismatch recognition)
  • High thermal stability of PNA–DNA or PNA–RNA duplexes
  • Resistance to enzymatic degradation

These properties make PNAs powerful tools in molecular diagnostics, antisense and antigene therapies, and other biomedical applications where strong, specific nucleic acid recognition is critical.

PNA vs. DNA: Backbone Structure and Binding

Backbone Structure Comparison
Binding Behavior
Applications
Molecular diagnostics
Gene targeting and editing research

Why PNAs Are Ideal for Diagnostic

Peptide Nucleic Acids (PNAs) are synthetic DNA mimics with a peptide-like backbone, enabling exceptionally strong and specific binding to complementary DNA or RNA. Their unique properties make them powerful tools for molecular diagnostics, offering unmatched sensitivity and accuracy. PNA offers several benefits in molecular detection:

Sequence-Specific Hybridization

PNAs bind to target sequences via Watson–Crick base pairing without the negative charge repulsion seen in DNA–DNA binding. Result: Higher stability and more reliable detection of target nucleic acids.

High Mismatch Discrimination

A single-base mismatch significantly reduces PNA binding stability, allowing precise differentiation between perfectly matched and mutated sequences. Result: Ideal for SNP genotyping and mutation detection .

PNA Clamping

PNAs can block DNA polymerase extension when bound to the target site. Result: Selective suppression of wild-type sequences in PCR, enabling mutant allele enrichment for rare mutation detection.

Signal-Generating PNA Probes

By attaching fluorescent dyes or quenchers, PNAs can produce a measurable signal upon hybridization.
Result: Used in real-time PCR, molecular beacons, and FISH assays for rapid and accurate detection.

In Situ Hybridization & Imaging

PNAs can penetrate cells and tissues when conjugated with delivery agents, binding directly to target nucleic acids. Result: Enables telomere length measurement, chromosome analysis, and pathogen identification in clinical samples.

Why PNAs Are Ideal for Diagnostics
  • Stronger binding than DNA or RNA probes
  • Greater specificity for detecting genetic variations
  • Resistant to enzymatic degradation, increasing assay robustness

Mechanisms of Action of PNA in Diagnostics

  1. Sequence-Specific Hybridization:
    PNAs bind to target sequences via Watson-Crick base pairing without charge repulsion.
  2. High Mismatch Discrimination:
    A single-base mismatch significantly reduces PNA binding stability.
  3. PNA Clamping:
    PNAs block DNA polymerase extension on the target site.
  4. Signal-Generating PNA Probes:
    PNAs produce a measurable signal upon hybridization.
  5. Signal-Generating PNA Probes:
    PNAs produce a measurable signal upon hybridization.
  6. In Situ Hybridization & Imaging:
    PNAs penetrate cells and tissues with delivery agents.

Key Advantages

Exceptional Stability

Resistant to nucleases and proteases for robust performance in complex matrices.

High Affinity & Specificity

Strong duplex/triplex formation with DNA/RNA; effective for clamping and discrimination.

Chemical Versatility

Neutral backbone supports diverse labels, linkers, and bioconjugations.

Applications

  • Molecular diagnostics & genotyping
  • Antisense/antigene research
  • qPCR & PCR clamping
  • FISH & in situ hybridization
  • Live-cell imaging probes
  • CRISPR off-target suppression (research)
  • Nanotechnology & sensors
  • PNA–peptide chimeras
  • PNA–oligo chimeras

Custom Synthesis Options

Parameter Options
Length 5–40 mer (longer upon review)
Purification HPLC (standard); PAGE optional
Labeling Positions N-terminus, C-terminus, internal
Scales 1 mg to gram quantities
Delivery Format Lyophilized; buffer on request
QC Package MALDI-TOF MS, analytical HPLC; optional Tm/hybridization

Popular Modifications & Linkers

Category Examples Common Uses
Fluorescent Dyes FAM, HEX, TET, TAMRA, ROX, Cy3, Cy5, ATTO series, Alexa Fluors Imaging, FISH/ISH, real-time assays
Affinity Tags Biotin (short/long spacer), Digoxigenin, DNP Capture, detection, pull-down
Reactive Handles Amine, Thiol, Azide, Alkyne (terminal/strained), Maleimide Bioconjugation via amide, thioether, or click reactions
Spacers / PEG Ahx, AEEA, miniPEG, PEG(n), C6/C12 alkyl linkers Reduce sterics; improve solubility/flexibility
Quenchers BHQ-1/2/3, Iowa Black, Eclipse Molecular beacons & quenched probes
Cleavables (on request) Disulfide, photocleavable (oNB), enzymatically cleavable spacers Triggered release & controlled activation

Conjugation Services

Type Linker Chemistry Applications
Peptide–PNA Amide coupling, maleimide–thiol, click (CuAAC/SPAAC) Cellular delivery, targeting peptides, CPPs
Oligo–PNA Click handles, heterobifunctional PEGs Hybrid probes, clamp constructs
Antibody–PNA SMCC/maleimide–NHS, site-selective strategies Immuno-detection, proximity ligation assays

Quality Assurance

  • MALDI-TOF MS for molecular weight confirmation
  • Analytical HPLC chromatogram and purity report
  • Optional: UV-melting (Tm), hybridization/functional assays
  • Documentation suitable for RUO; GLP/cGMP support upon request

Typical Turnaround

Standard labeled PNA: 2–3 weeks from order confirmation. Complex chimeras or antibody conjugations may require additional time.

Lead time is sequence- and modification-dependent; rush options may be available.

How to Order

  1. Provide your sequence (or request design assistance).
  2. Select labels, linkers, and conjugations.
  3. Choose scale, purification, and any optional tests.
  4. Receive a same-day quote and projected timeline.

Have an RFP or multi-sequence panel? Attach it in the form below.

Specifications Checklis

  • Sequence (5'→3'), length, and any degeneracy
  • Desired modifications & positions
  • Conjugation partner (if any) and linker preference
  • Scale, purification, and QC requirements

PNA Technology & Benefits

What are PNAs? Peptide Nucleic Acids (PNAs) are DNA mimics in which the sugar–phosphate backbone is replaced by a neutral, peptide-like chain of N-(2-aminoethyl)-glycine units linked by amide bonds. Bases (A, C, G, T) attach via methylene-carbonyl linkages, preserving Watson–Crick pairing. The uncharged backbone removes electrostatic repulsion, yielding fast, high-affinity, and highly specific hybridization to complementary DNA or RNA.

How PNAs Work

  • Strong duplex formation: Neutral backbones enable tighter PNA•DNA/RNA duplexes and faster on-rates, even at lower ionic strength.
  • Sharp mismatch discrimination: Single-base mismatches destabilize PNA hybrids more than DNA/DNA, enabling SNP-level specificity.
  • PNA clamp effect: Polymerases cannot extend through a bound PNA, allowing selective suppression of wild-type alleles in PCR for rare-mutation detection.
  • Enzymatic stability: PNAs resist nucleases and proteases, supporting robust performance in complex samples.
  • Low-background FISH: Short, high-affinity probes reduce reannealing of genomic strands under low-salt conditions, improving signal-to-noise.

Design Principles

  • Length: Start with ~12–18-mers; tune to target Tm and assay stringency.
  • Buffer/stringency: PNAs tolerate lower salt—use to suppress genomic reannealing (useful for repetitive targets).
  • Solubility & handling: Add lysine/PEG spacers or AEEA to improve solubility for longer or dye-rich probes.
  • Placement: For clamp PCR, cover the wild-type site; for FISH, avoid strong secondary structures and add spacers to reduce quenching.
  • Labeling: Compatible with fluorescein, rhodamine, cyanine, Alexa Fluor®, quenchers, and biotin for capture.

Benefits at a Glance

  • High affinity & thermal stability vs. DNA probes
  • Exceptional specificity (single-nucleotide discrimination)
  • Fast hybridization and salt independence
  • Lower background in FISH/ISH assays
  • Enzymatic resistance for durable performance
  • Shorter probes with equal or better signal
  • Versatile labeling & conjugation for diagnostics and capture

PNAs bind strongly—overly long designs can overshoot Tm. We optimize probe length, spacers, and labels to hit your assay window with clean specificity.

Use Case Why PNA Helps Typical Setup
Rare-mutation detection (PNA clamp PCR) Blocks wild-type extension; enriches mutant signal PNA clamp overlapping WT site; mutation-specific primers/probe; qPCR or ddPCR readout
FISH/ISH (Telomere, Centromere, Pathogens) High affinity + low reannealing → strong S/N 12–18-mer dye-labeled PNA; low-salt hybridization; optional formamide; spacers to limit quenching
SNP genotyping Single-base mismatch discrimination Short PNA probe across variant; melt-curve, beacon, or real-time probe formats
Capture & pull-down Tight binding and nuclease resistance 5′ biotin-PNA with PEG spacer; streptavidin capture; stringent washes
In situ pathogen ID Short, specific probes in complex matrices Dye-labeled PNA panels targeting species markers; multiplex imaging
Optimize My PNA Design

FAQ

What sequence lengths are recommended for PNAs?

Most applications use 10–20-mers. Short (8–12) for clamping/high specificity, longer for challenging targets. We can advise based on your assay.

Can PNAs be used in live-cell assays?

Yes. Delivery often benefits from peptide conjugation (e.g., CPPs) or optimized spacers/PEG to improve uptake and reduce steric hindrance.

What purity do you provide?

HPLC-purified PNAs are standard. PAGE purification is available on request. Each lot ships with MALDI-TOF and analytical HPLC reports.

Do you offer GMP production?

Yes. We support RUO through cGMP with appropriate documentation and QA oversight. Please include your regulatory needs in the request.

Speak to a Scientist

Complete the form to receive a tailored quote. Your request will be emailed to info@biosyn.com and logged to your CRM endpoint (configure below).

Full Name *
Email *
Company / Institution *
Phone *
PNA Sequence(s)
Labels / Linkers / Conjugations
Scale *
Purification
Notes / Requirements

By submitting, you agree to be contacted regarding your request.

Why Choose Bio-Synthesis

Trusted by biotech leaders worldwide for over 40+ years of delivering high quality, fast and scalable synthetic biology solutions.