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Maxam-Gilbert Sequencing

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In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
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Updated: Jan 9, 2026

Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids

Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids

Published on: September 21, 2017

7-Methylguanine With a Cyclopentane Backbone: A Bright Combination for a FIT-PNA RNA Sensor.

Salam Maree¹, Pinaki Chanda¹, Sheethal Thomas Mannully¹

¹The Institute for Drug Research, The School of Pharmacy, The Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, Israel.

British Journal of Biomedical Science

|December 8, 2025

Summary

Chemically modified FIT-PNAs (forced intercalation-Peptide Nucleic Acids) show significantly enhanced brightness and specificity for detecting cancer-associated long noncoding RNA (lncRNA CCTA-1) in cells.

Keywords:

BisQ FIT-PNA RNA biosensors cpG+molecular simulations

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Kinetic Screening of Nuclease Activity using Nucleic Acid Probes

Kinetic Screening of Nuclease Activity using Nucleic Acid Probes

Published on: November 1, 2019

Area of Science:

Chemical Biology
Molecular Diagnostics
Biotechnology

Background:

FIT-PNAs (forced intercalation-Peptide Nucleic Acids) offer potential as RNA sensors due to fluorescence enhancement upon hybridization.
Existing FIT-PNAs require optimization for increased brightness and specificity in biological applications.

Purpose of the Study:

To develop a novel chemical modification strategy for FIT-PNAs to enhance their fluorescence and specificity for RNA detection.
To investigate the impact of modifying the guanine base adjacent to the fluorophore on FIT-PNA performance.

Main Methods:

Synthesized FIT-PNAs with chemically modified guanine bases (G+, cpG, cpG+) adjacent to the BisQ surrogate base.
Tested FIT-PNA performance using synthetic RNA and DNA targets, including sequences with mismatches.
Evaluated FIT-PNA fluorescence and specificity in ovarian cancer cells overexpressing CCAT1 using FACS and confocal microscopy.
Employed molecular simulations to correlate experimental fluorescence data with molecular interactions.

Main Results:

The cpG+ modified FIT-PNA exhibited a two-fold increase in brightness compared to the unmodified guanine base.
Modified FIT-PNAs showed increased fluorescence with RNA mismatches, unlike synthetic DNA.
The cpG+ modified FIT-PNA successfully generated a bright fluorescent signal in ovarian cancer cells, enabling visualization of CCAT1.
Molecular simulations supported the experimental findings regarding fluorescence and mismatch behavior.

Conclusions:

Chemical modification of the guanine base adjacent to the fluorophore significantly enhances FIT-PNA brightness and RNA specificity.
The cpG+ modification represents a promising advancement for developing highly sensitive and specific RNA sensors for biomedical applications.
This strategy holds potential for improved molecular diagnostics and targeted therapies in diseases like cancer.