Performing encapsulation of dsDNA and a polymerase chain reaction (PCR) inside nanocontainers using the inverse miniemulsion process.
We report the encapsulation of dsDNA molecules with a defined number of base pairs (476 bp and 790 bp) and their subsequent amplification by polymerase chain reaction (PCR) inside nanosized polymeric capsules/droplets. In the first set of experiments, the dsDNA template and PCR reagents were encapsulated in crosslinked potato starch using the inverse (water-in-oil) miniemulsion technique. After redispersion of the capsules in a water-surfactant mixture, PCR was performed inside the crosslinked starch nanocapsules. In the second set of experiments, the PCR was performed inside the aqueous nanodroplets before capsule formation, and then each miniemulsion droplet was covered with a polybutylcyanoacrylate (PBCA) shell which was formed through anionic polymerization directly at the droplet interface. The PCR efficiency was quantitatively evaluated by fluorescence spectroscopy, using a DNA-specific dye called SYBR® Green which intercalates between the base pairs of the dsDNA.
Max Planck Institute for Polymer Research, Mainz, Germany.
Grit
G
Baier
Anna
A
Musyanovych
Volker
V
Mailänder
Katharina
K
Landfester
D003432:Cross-Linking Reagents / Q000737:chemistry; D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry; D004655:Emulsions; D004659:Enbucrilate / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D018014:Gene Transfer Techniques; D007364:Intercalating Agents / Q000737:chemistry; D053769:Nanocapsules; D036103:Nanotechnology; D009690:Nucleic Acid Conformation; D010316:Particle Size; D016133:Polymerase Chain Reaction; D011108:Polymers / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013213:Starch / Q000737:chemistry; D013501:Surface-Active Agents / Q000737:chemistry
D003432:Cross-Linking Reagents; D004655:Emulsions; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D053769:Nanocapsules; D011108:Polymers; D013501:Surface-Active Agents; D013213:Starch; D004247:DNA; D004659:Enbucrilate