Tools and Techniques

Biochemistry

Biochemical techniques, including PCR, in vitro transcription, translation and methods of purification, such as gel electrophoresis and HPLC will be used for the synthesis and isolation of unique biomolecules utilized in the group. In addition to these various biochemical approaches, several molecular biology techniques will be utilized for expressing plasmids in cells and for the intracellular delivery and uptake of nanomaterials.

                                RNA structure                    gel jpeg

Above: Left, an example of a hammerhead ribozyme*, a catalytic RNA structure capable of cleaving other RNA.  New functional RNA structures will be evolved through in vitro selection.  Right, fluorescently labeled RNA-nanoparticle conjugates run through an agarose gel.

Chemical and Materials Synthesis

New chemical attachment strategies will be developed which will allow nucleic acids and peptides to be successfully interfaced with inorganic and organic nanomaterials for various applications.

          probe jpeg

Above: Phosphorus 31 NMR showing the synthesis of an amino acid (alanine) phosphoramidite useful for adding protein residues to nucleic acids.

Below: TEM image of CdS quantum dots interfaced with Pt rich nanoparticles.  EDS spectrum indicates relative elemental amounts.

               eds w tem

Nanoscale Materials Characterization

Several forms of microscopy, including scanning electron microscopy (SEM), transmission electron microscopy (TEM), and atomic force microscopy (AFM) will be utilized for characterizing the biomaterials designed within the lab. Compositional analysis including Inductively Coupled Plasma Mass Spectroscopy (ICP-MS) and Energy Dispersive Spectroscopy (EDS) will be used for confirming the structure and make up of nanomaterials interfaced with nucleic acids and peptides. In addition, fluorescence spectroscopy techniques will be utilized for characterizing the photophysical properties of the materials generated and for designing assays that can probe the interactions between nucleic acid nanoparticle conjugates and their intended targets.

Below: TEM image of Pt nanoparticles.

Pt NPs jpeg

*Martick M.; Scott, W. G. Cell, 2006, 126, 309-320.