摘要:
A process for the qualitative and quantitative determination of at least one in vitro amplified nucleic acid in a sealed reaction chamber,wherein during or subsequent to the amplification of the nucleic acid at least one substance (probe) is present which interacts with the nucleic acid to be detected;wherein spectroscopically measurable parameters of said substance (probe) are subject to variation, creating a measurable signal;wherein the sample to be measured is exposed to the action of a gradient capable of at least partially denaturing nucleic acids;with detection of the measurable parameter undergoing variation through the action of the gradient; andthe entire amplification reaction, including qualitative and quantitative determination, may be carried out in a sealed reaction chamber (measuring compartment) without intermittent opening,permitting to automatically operate the diagnostic method of DNA and RNA amplification in qualitative and quantitative fashion on large series of samples.
摘要:
The invention relates to a process for the separation and detection of components of a mixture of materials by temperature gradient gel electrophoresis, wherein either a spatial temperature gradient is built up by spatially separated temperature levels, or a time temperature gradient, or a temperature gradient is built up by combination of spatial and timewise temperature gradient. The temperature levels for building up the spatial temperature gradient are adjusted by controllable heating or cooling devices. To build up the time temperature gradient, the temperature level at each point of the separation path within the separation medium may be optionally adjusted time-dependently by means of controllable heating or cooling devices. There is described a device for performing the process with controllable heating or cooling devices to build up temperature gradients, a hollow body arranged between the temperature levels which contains the medium used for separation, and a thermostat jacket enclosing the hollow body.
摘要:
A Method for identifying one or a small number of molecules, especially in a dilution of ≦1 μM, using laser excited FCS with measuring times ≦500 ms and short diffusion paths of the molecules to be analyzed, wherein the measurement is performed in small volume units of preferably ≦10−14 l, by determining material-specific parameters which are determined by luminescence measurements of molecules to be examined.The device which can be preferably used for performing the method according to the invention is a per se known system of microscope optics for laser focusing for fluorescence excitation in a small measuring compartment of a very diluted solution and for imaging the emitted light in the subsequent measurement through confocal imaging wherein at least one system of optics with high numerical aperture of preferably ≧1.2 N.A. is employed, the light quantity is limited by a confocally arranged pinhole aperture in the object plane behind the microscope objective, and the measuring compartment is positioned at a distance of between 0 and 1000 μm from the observation objective.
摘要:
The process of generating new biopolymers with improved characteristics, assisted by polymerases, is characterized in that at least one cycle of the steps described below, will be performed in a series of parallel preparations which are to be compared. Sequences of nucleic acids or, a mixture of similar sequences of nucleic acids in the distribution of mutants of a quasi-species, are subjected, in the region of the error threshold, to limited mutagenesis. These mixtures are replicated under simultaneous conditions and/or in succession. The mixtures of the so product nucleic acids are compartmentalized by segregation, and thereafter selected by a selection system which reflects the characteristics of interest of the nucleic acid sequence itself or, indirectly, via its translation product.
摘要:
A process for instabilizing viral quasi-species-distributions under avoidance of resistance phenomena by replication of the nucleic acids of the viruses present in the quasi-species-distribution by of a defective replication system, a) whereby the defective replication system has a rate of misincorporation for nucleotides above the rate of misincorporation of the viral wild-type-replication system and, whereby the viruses are replicated by the replication system having the higher rate of misincorporation at least as effectively as it is done by the replication system of the wild-type virus, b) and/or negative influence of the replication of the consensus-sequence (nucleic acid sequence of the wild-type virus) in relation to other replicatable nucleic acids.
摘要:
A process for instabilizing viral quasi-species-distributions under avoidance of resistance phenomena by replication of the nucleic acids of the viruses present in the quasi-species-distribution by of a defective replication system, a) whereby the defective replication system has a rate of misincorporation for nucleotides above the rate of misincorporation of the viral wild-type-replication system and, whereby the viruses are replicated by the replication system having the higher rate of misincorporation at least as effectively as it is done by the replication system of the wild-type virus, b) and/or negative influence of the replication of the consensus-sequence (nucleic acid sequence of the wild-type virus) in relation to other replicatable nucleic acids.
摘要:
Described is a support material that can simultaneously bind both genotypic and phenotypic substances. The respective areas, which can have surface-modifying matter such as anionic exchangers and/or affinity ligands, simultaneously bind, for example, nucleic acids and proteins or peptides. The support materials described can be used in processes for evolutive optimization of biopolymers, wherein genotype and phenotype can be bound at the same time so as to furnish them for further analysis.
摘要:
The method according to the invention permits a selected withdrawal of one or a few molecularly disperse or cellular components of a system, such as molecules, molecular complexes, vesicles, micelles, cells, optionally together with an associated volume element V having a size of 10−9 l≧V≧10−18 l from a larger sample volume. The selected transfer of the sought component to another environment is effected by defining the space and time of withdrawal by means of a signal correlating with the small component to be withdrawn. The method is particularly useful for the withdrawal of non-abundant components the existence of which can be detected in a preceding step by a scanning process. The method is also useful for the withdrawal of per se unidentified components.
摘要:
The method according to the invention permits a selected withdrawal of one or a few molecularly disperse or cellular components of a system, such as molecules, molecular complexes, vesicles, micelles, cells, optionally together with an associated volume element V having a size of 10−9 l≧V≧10−18 l from a larger sample volume. The selected transfer of the sought component to another environment is effected by defining the space and time of withdrawal by means of a signal correlating with the small component to be withdrawn. The method is particularly useful for the withdrawal of non-abundant components the existence of which can be detected in a preceding step by a scanning process. The method is also useful for the withdrawal of per se unidentified components.
摘要:
A Method for identifying one or a small number of molecules, especially in a dilution of ≦1 μM, using laser excited FCS with measuring times ≦500 ms and short diffusion paths of the molecules to be analyzed, wherein the measurement is performed in small volume units of preferably ≦10−14 l, by determining material-specific parameters which are determined by luminescence measurements of molecules to be examined. The device which can be preferably used for performing the method according to the invention is a per se known system of microscope optics for laser focusing for fluorescence excitation in a small measuring compartment of a very diluted solution and for imaging the emitted light in the subsequent measurement through confocal imaging wherein at least one system of optics with high numerical aperture of preferably ≧1.2 N.A. is employed, the light quantity is limited by a confocally arranged pinhole aperture in the object plane behind the microscope objective, and the measuring compartment is positioned at a distance of between 0 and 1000 μm from the observation objective.