Graphene Oxide/Nitrocellulose Non-Covalent Hybrid as Solid Phase for Oligo-DNA Extraction from Complex Medium

Area of Science:

• Materials Science
• Biotechnology
• Analytical Chemistry

Background:

• Oligonucleotide extraction from complex biological matrices is crucial for molecular diagnostics and research.
• Existing methods can be inefficient or require complex procedures.
• Development of novel, high-performance materials for nucleic acid purification is an ongoing need.

Purpose of the Study:

• To develop and characterize a novel nitrocellulose-graphene oxide (NC-GO) hybrid membrane for efficient oligonucleotide extraction.
• To evaluate the performance of the NC-GO hybrid membrane in purifying short oligonucleotides (<50 nucleotides) from various complex solutions.
• To optimize the extraction conditions, including incubation time and medium composition.

Main Methods:

• Preparation of a nitrocellulose (NC) membrane non-covalently modified with graphene oxide (GO) microparticles.
• Characterization of the NC-GO hybrid membrane using Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM).
• Wettability assessment via water contact angle measurements.
• Testing the extraction efficiency of oligonucleotides from aqueous medium, α-Minimum Essential Medium (αMEM), and αMEM with fetal bovine serum (FBS) over different time periods (30, 45, 60 min).
• Desorption of extracted oligonucleotides using Tris-HCl buffer (pH 8.0) and quantification via fluorescence emission.

Main Results:

• FTIR and SEM confirmed successful, uniform modification of the NC membrane with GO, exhibiting a spider web morphology.
• The NC-GO hybrid membrane showed slightly reduced hydrophilicity (contact angle 26.7°) compared to the bare NC membrane (15°).
• Optimal oligonucleotide extraction was achieved after 60 min incubation in αMEM, yielding the highest fluorescence emission (294 r.f.u.), corresponding to approximately 330-370 pg (7%) of total oligo-DNA.
• The NC-GO hybrid membrane demonstrated effective purification of short oligonucleotides from complex biological solutions.

Conclusions:

• The developed nitrocellulose-graphene oxide hybrid membrane is a promising material for the efficient and effortless purification of short oligonucleotides.
• This method offers a simplified approach for isolating target nucleic acid sequences from challenging sample matrices.
• The NC-GO hybrid membrane represents a valuable tool for applications in molecular biology, diagnostics, and synthetic biology.