Irreversible trapping of DNA during crossed-field gel electrophoresis.

Using an original protocol with a rotating gel electrophoresis apparatus, it is shown that duplex DNA undergoing crossed-field electrophoresis in agarose gets trapped in the gel when the field is increased above a threshold value which decreases with the chain length and depends on the angle between the fields in a non-monotonous manner. This trapping is irreversible, i.e. once trapped at a high field strength, chains are unable to resume their motion when the field is returned to a lower value at which they moved prior to trapping. A model of trapping by "tight knots" is proposed. It predicts a trapping threshold proportional to the inverse square of the electric field, in qualitative agreement with the data. The implications of our results for the separation of large DNA molecules are discussed.     

PHOTOOXIDATION OF CHLOROPHYLL AND PHEOPHYTIN. QUENCHING OF SINGLET OXYGEN AND INFLUENCE OF THE MICELLAR STRUCTURE

Abstract— When chlorophyll(Chl) and pheophytin(Phn) are irradiated in Triton X-100 water binary solvents, singlet oxygen is formed in the medium in a higher yield for Phn than for Chi. Chlorophyll shows an irreversible photooxidation reaction and a chemical oxidation reaction when 1,3-diphenyliso-benzofuran (DPBF) is added to the solution. During the chemical oxidation, Chi is destroyed by an oxidizing agent that is a reaction product of the endoperoxide formed in the medium by the addition of singlet oxygen to DPBF. This reaction depends on the structure of the medium and has some characteristics of an oxidation by hydroxyl radicals. The highest yield is obtained with the micellar structure. Chlorophyll and Phn are readily oxidized by hydroxyl radicals generated using the Fenton reagent. This suggests that in the presence of Triton X-100, the Mg²⁺ ion of a Chi molecule plays a key role in the irreversible oxidation of the pigment.     

Self-assembled magnetic nanowires made irreversible by polymer bridging

In this letter, we investigate the mechanism of formation of a recently discovered new type of colloid, irreversible flexible chains of magnetic particles. The chain formation mechanism is based on magnetically induced bridging by adsorbed polymers, and we investigate here the associated phase diagrams, considering both thermodynamic and kinetic aspects. This phase diagram is the consequence of a balance between entropic repulsion between polymer layers at the particles surfaces, depletion forces pushing the particles together, and a short-range attractive force developing when polymers can bridge two particles. We end up with a very simple protocol allowing the formation of long, extremely regular chains, which can find numerous applications in chemistry and biology. The perspectives for the development of a new field of "macrocolloidal chemistry" are discussed.     

New block-copolymer thermoassociating matrices for DNA sequencing: effect of molecular structure on rheology and resolution

A new family of matrices for DNA sequencing by capillary electrophoresis is presented. These matrices combine easy injection with high sieving performances, due to thermal switching between a low and a high viscosity state through a modest increase in temperature (approximately 20 degrees C). They are constructed from a hydrophilic polymer backbone with grafted lower critical solution temperature (LCST) side chains. The comb-like LCST copolymers are characterized in terms of size of the polymer backbone, the size of LCST side chains and the grafting densities. The dependance of rheological behavior and electrophoretic performance of these copolymers is correlated with their microstructure. Without complete optimization, a resolution of order 0.5, corresponding to a very reasonable limit for read length with current base calling softwares, could be achieved for segments around 800 bases differing by 1 base in less than one hour in a commercial ABI 310 apparatus.     

Comportement non lorentzien de la raie Rayleigh dépolarisée dans quelques liquides. Discussion des théories ‘à trois variables’

Nous considérons en général le spectre Rayleigh dépolarisé des liquides simples constitués de molécules anisotropes comme la superposition d'une raie lorentzienne de réorientation, comportant une structure fine dûe au couplage entre orientation et modes hydrodynamiques, et d'une aile lointaine d'origine collisionnelle. Par une analyse détaillée des spectres d'un certain nombre de liquides constitués de molécules isotropes (CCl₄) ou anisotropes (benzène, pyridine, cyanopropyne, 2-butyne, iodopropyne), cet article met en évidence pour ces derniers une quatrième ‘composante’, bien représentée par une lorentzienne de demi-largeur comprise entre 3 et 10 cm^-1. Cette deuxième lorentzienne peut représenter jusqu'à 30 pour cent de l'intensité totale du spectre. Nous montrons qu'elle ne peut être dûe à une simple anisotropie réorientationnelle décrite par la variable tensorielle i. Elle implique donc l'existence d'une seconde variable tensorielle d'anisotropie locale g, attribuée à l'anisotropie d'ordre translationnel.

Les théories dites ‘à trois variables’, supposant g directe (primaire) ou indirecte (secondaire), sont comparées à la lumière de ces résultats expérimentaux, sans qu'il soit possible de trancher définitivement cette question. En particulier, nous montrons que dans le cas le plus général — g à la fois directe et indirecte-il existe plus de paramètres théoriques que de paramètres déductibles de l'expérience.     

Tube relaxation: A necessary concept in the dynamics of strained polymers

A theory for the relaxation of large strains in polymer melts is outlined. It is based on the Doi–Edwards slip-link model and the new concept of tube relaxation. Self-consistency makes this concept necessary when polymer melts are concerned. The discrepancy with previous non-self-consistent theories is not negligible and should be easy to observe experimentally. Special attention is given to the overall size and to the mean orientation of a labeled N-chain in a step-strained melt of P-chains. Using semiquantitative arguments, we predict different regimes depending on the respective values of N and P, and propose approximate evolution equations for each case. These equations may be used to design and interpret a variety of fluorescence, infrared, or NMR polarization measurements, as well as small-angle neutron scattering experiments.     

Droplet fusion by alternating current (AC) field electrocoalescence in microchannels

We present a system for the electrocoalescence of microfluidic droplets immersed in an immiscible solvent, where the undeformed droplet diameters are comparable to the channel diameter. The electrodes are not in direct contact with the carrier liquid or the droplets, thereby minimizing the risk of cross-contamination between different coalescence events. Results are presented for the coalescence of buffered aqueous droplets in both quiescent and flowing fluorocarbon streams, and on-flight coalescence is demonstrated. The capillary-based system presented here is readily amenable to further miniaturization to any lab-on-a-chip application where the conductivity of the droplets is much greater than the conductivity of the stream containing them, and should aid in the further application of droplet microreactors to biological analyses.