Precise mass measurement of a double-stranded 500 base-pair (309 kDa) polymerase chain reaction product by negative ion electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry

Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICRMS) has been used to determine the mass of a double-stranded 500 base-pair (bp) polymerase chain reaction (PCR) product with an average theoretical mass of the blunt-ended (i.e. unadenylated) species of 308 859.35 Da. The PCR product was generated from the linearized bacteriophage Lambda genome which is a double-stranded template. Utilization of ethanol precipitation in tandem with a rapid microdialysis step to purify and desalt the PCR product was crucial to obtain a precise mass measurement. The PCR product (0.8 pmol/μL) was electrosprayed from a solution containing 75% acetonitrile, 25 mM piperidine, and 25 mM imidazole and was infused at a rate of 200 nL/min. The average molecular mass and the corresponding precision were determined using the charge-states ranging from 172 to 235 net negative charges. The experimental mass and corresponding precision (reported as the 95% confidence interval of the mean) was 309 406 +/- 27 Da (87 ppm). The mass accuracy was compromised due to the fact that the PCR generates multiple products when using Taq polymerase due to the non-template directed 3'-adenylation. This results in a mixture of three PCR products with nearly identical mass (i.e. blunt-ended, mono-adenylated and di-adenylated) with unknown relative abundances that were not resolved in the spectrum. Thus, the experimental mass will be a weighted average of the three species which, under our experimental conditions, reflects a nearly equal concentration of the mono- and di-adenylated species. This report demonstrates that precise mass measurements of PCR products up to 309 kDa (500 bp) can be routinely obtained by ESI-FTICR requiring low femtomole amounts. Copyright 1999 John Wiley & Sons, Ltd.     

Mach reflection in detonations propagating through a gas with a concentration gradient

In accident scenarios where detonations can occur a concentration gradient constitutes a more realistic initial condition than a perfectly homogeneous mixture. In this paper, the influence of a concentration gradient on detonation front shape, detonation instabilities and pressure distribution is studied. First, a simple method to determine the front shape from a given fuel distribution is presented. It is based on Huygens’ principle and includes a correction to satisfy the boundary conditions on the enclosing walls. Next, the presented highly resolved Euler computations demonstrate the influence of a concentration gradient on detonation instabilities. In configurations with a strong concentration gradient, Mach reflection occurs and leads to an asymmetric pressure load on the enclosing geometry. In this case, the impulse on the wall is higher than in configurations with homogeneous fuel distribution, although the fuel content is much lower.     

Enhanced nanoparticle detection with liquid droplet resonators

Whispering gallery mode particle sensing experiments are commonly performed with solid resonators, whereby the sensing volume is limited to the weak evanescent tail of the mode near the resonator surface. In this work we discuss in detail the sensitivity enhancements achievable in liquid droplet resonators wherein the stronger internal fields and convenient means of particle delivery can be exploited. Asymptotic formulae are derived for the relative resonance shift, line broadening and mode splitting of TE and TM modes in liquid droplet resonators. As a corollary the relative fraction of internal and external mode energy follows, which is shown to govern achievable sensitivity enhancements of solute concentration measurements in droplet sensors. Experimental measurements of nanoparticle concentration based on whispering gallery mode resonance broadening are also presented.     

Intermolecular hybridization governs molecular electrical doping

Current models for molecular electrical doping of organic semiconductors are found to be at odds with other well-established concepts in that field, like polaron formation. Addressing these inconsistencies for prototypical systems, we present experimental and theoretical evidence for intermolecular hybridization of organic semiconductor and dopant frontier molecular orbitals. Common doping-related observations are attributed to this phenomenon, and controlling the degree of hybridization emerges as a strategy for overcoming the present limitations in the yield of doping-induced charge carriers.     

A model-theoretic characterization of constant-depth arithmetic circuits

We study the class ${\mathrm{\#}\text{AC}}^0$ of functions computed by constant-depth polynomial-size arithmetic circuits of unbounded fan-in addition and multiplication gates. No model-theoretic characterization for arithmetic circuit classes is known so far. Inspired by Immerman's characterization of the Boolean circuit class ${\text{AC}}^0$, we remedy this situation and develop such a characterization of ${\mathrm{\#}\text{AC}}^0$. Our characterization can be interpreted as follows: Functions in ${\mathrm{\#}\text{AC}}^0$ are exactly those functions counting winning strategies in first-order model checking games. A consequence of our results is a new model-theoretic characterization of ${\text{TC}}^0$, the class of languages accepted by constant-depth polynomial-size majority circuits.