Reversible DNA encapsulation in silica to produce ROS-resistant and heat-resistant synthetic DNA 'fossils'

• Published in: Nature protocols Vol. 8; no. 12; pp. 2440 - 2448
• Main Authors: Paunescu, Daniela, Puddu, Michela, Soellner, Justus O B, Stoessel, Philipp R, Grass, Robert N
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.12.2013; Nature Publishing Group
• Subjects: 631/1647/350/2093; 631/1647/666/2263; 631/92/147; 639/638/45/2783; Acids; Ammonium; Analytical Chemistry; Biological Techniques; Computational Biology/Bioinformatics; Deoxyribonucleic acid; DNA; DNA - chemistry; DNA - ultrastructure; DNA, Single-Stranded - chemistry; Electrophoresis; Etching; Fluorides; Fossils; Genetic research; Heat; High temperature; Hot Temperature; Hydrogen fluoride; Life Sciences; Microarrays; Microencapsulation; Microscopy, Electron, Transmission; Nanoparticles; Nucleic acids; Organic Chemistry; Plasmids - genetics; Preservation, Biological - methods; Properties; Protocol; Reactive oxygen species; Reactive Oxygen Species - chemistry; Silica; Silicon Dioxide - chemistry; Surface Properties
• ISSN: 1754-2189; 1750-2799; 1750-2799
• DOI: 10.1038/nprot.2013.154

This protocol describes a method for encapsulating DNA into amorphous silica (glass) spheres, mimicking the protection of nucleic acids within ancient fossils. In this approach, DNA encapsulation is achieved after the ammonium functionalization of silica nanoparticles. Within the glass spheres, the nucleic acid molecules are hermetically sealed and protected from chemical attack, thereby withstanding high temperatures and aggressive radical oxygen species (ROS). The encapsulates can be used as inert taggants to trace chemical and biological entities. The present protocol is applicable to short double-stranded (ds) and single-stranded (ss) DNA fragments, genomic DNA and plasmids. The nucleic acids can be recovered from the glass spheres without harm by using fluoride-containing buffered oxide etch solutions. Special emphasis is placed in this protocol on the safe handling of these buffered hydrogen fluoride solutions. After dissolution of the spheres and subsequent purification, the nucleic acids can be analyzed by standard techniques (gel electrophoresis, quantitative PCR (qPCR) and sequencing). The protocol requires 6 d for completion with a total hands-on time of 4 h.