Toward a microchip-based solid-phase extraction method for isolation of nucleic acids

A silica-based solid-phase extraction system suitable for incorporation into a microchip platform (nu-total analytical system; nu-TAS) would find utility in a variety of genetic analysis protocols, including DNA sequencing. The extraction procedure utilized is based on adsorption of the DNA onto bare silica. The procedure involves three steps: (i) DNA adsorption in the presence of a chaotropic salt, (ii) removal of contaminants with an alcohol/water solution, and (iii) elution of the adsorbed DNA in a small volume of buffer suitable for polymerase chain reaction (PCR) amplification. Multiple approaches for incorporation of this protocol into a microchip were examined with regard to extraction efficiency, reproducibility, stability, and the potential to provide PCR-amplifiable DNA. These included packing microchannels with silica beads only, generating a continuous silica network via sol-gel chemistry, and combinations of these. The optimal approach was found to involve immobilizing silica beads packed into the channel using a sol-gel network. This method allowed for successful extraction and elution of nanogram quantities of DNA in less than 25 min, with the DNA obtained in the elution buffer fraction. Evaluation of the eluted DNA indicated that it was of suitable quality for subsequent amplification by PCR.     

Enzymatic synthesis of chiral organophosphothioates from prochiral precursors

The phosphotriesterase from Pseudomonas diminuta has been shown to selectively cleave the pro-R p-nitrophenolate substituent from bis-p-nitrophenyl alkyl phosphothioate esters. When the alkyl substituent is methyl, ethyl, or isopropyl the enantiomeric excess of the product is >/=99%. Manipulation of the active site through mutagenesis has enabled the preparation of protein variants that preferentially hydrolyze the pro-S substituent of the target substrates. This methodology thus permits the preparation of chiral products from prochiral precursors.     

A high-speed high-sensitivity acoustic cell for in-line continuous monitoring of MOCVD precursor gases

We describe a continuous wave resonant acoustic sensor that has been optimized as a very sensitive in-line monitor for measuring the composition of precursor gases used in MOCVD processes. The precursor/carrier gas mixtures flow through a compact stainless steel acoustic chamber that is isolated from the acoustic transducers by a set of metallic diaphragms. The sensor has been successfully operated at supply line pressures from atmosphere down to 50 Torr with gas flow rates of up to 1600 sccm. The accuracy of the speed of sound measurement for hydrogen gas is better than 0.005%, even in a high noise and low pressure environment. Hydrogen, as well as nitrogen or argon carrier gases, are accommodated within the instrument's 1–5 kHz working frequency range. The instrument's sensitivity and stability are demonstrated with the laboratory data. Measurements of the dynamic response characteristics of the metalorganic bubbler lines at low pressure are also be presented. Application of the cell is general, encompassing any of the metalorganic and hydride materials typically used in MOCVD processes.     

Synthesis and thermal behavior of dimethyl scandium complexes featuring anilido-phosphinimine ancillary ligands

A family of N,N donor ligands [1-(NHAr)-2-(PR₂NAr′)C₆H₄] (1a-d; Ar = 2,6-ⁱPr₂-C₆H₃, R = Me, Ph, Ar′ = 2,4,6-Me₃-C₆H₂, 2-ⁱPr-C₆H₄, 2,6-ⁱPr₂-C₆H₃) has been prepared and fully characterized by multinuclear NMR spectroscopy and X-ray crystallography. Lithiation of the N-H unit and subsequent salt metathesis protocols with ScCl₃THF₃ provides an avenue to organometallic scandium complexes. The resultant base-free monomeric dichlorides LScCl₂, 3a-d, have been fully characterized by NMR spectroscopy as well as X-ray crystallography (3a,c,d). Alkylation of the dichlorides using LiMe results in clean formation of dialkyl complexes LScMe₂4a-c. Thermolysis of these materials under argon and hydrogen leads to decomposition products as a result of C-H activation of the ligand. Analysis of these results provides a qualitative assessment of the metalative resistance of each ligand framework.     

Thermal decomposition of poly(butylene terephthalate)

Detailed results of the overall thermal degradation of poly(butylene terephthalate) are reported. Laser microprobe analysis and dynamic mass spectrometric techniques were used to identify the primary volatile degradation products and initial pyrolysis reactions that control polymer degradation. A complex multistage decomposition mechanism was observed which involves two major reaction pathways. Initial degradation occurs by an ionic decomposition process that results in the evolution of tetrahydrofuran. This is followed by concerted ester pyrolysis reactions that involve an intermediate cyclic transition state and yield 1,3-butadiene. Simultaneous decarboxylation reactions occur in both decomposition regimes. Finally, the latter stages of polymer decomposition were characterized by evolution of CO and complex aromatic species such as toluene, benzoic acid, and terephthalic acid. Activation energies of formation for the main pyrolysis products were determined from the dynamic measurements of the major ion species and indicate values of E = 27.9 kcal/mole for the production of tetrahydrofuran and E = 49.7 kcal/mole for the production of butadiene.     

Synthesis of Nitrated Analogs of Distamycin and their DNA Cleaving and Protecting Activities under UV Irradiation

Two nitrated analogs of distamycin were synthesized by a novel haloform reaction and the DCC/HOBT coupling reaction in a convenient and effective way with out amino group protection. Our results have demonstrated that these polyamides show DNA cleaving activity under long UV (LUV 365 nm) irradiation. Furthermore, it is an unexpected discovery that these analogs can prevent DNA dam age under short UV (SUV 302 nm).