30 results about "Enzymatic amplification" patented technology

A method for multiple and simultaneously detecting and quantifying of Listeria spp, Staphylococcus aureus, Campylobacter jejuni, and Escherichia coli 0157:H7, by extracting DNA from a sample; preparing a reaction mixture for enzymatic amplification of the extracted DNA and sets of pairs of oligonucleotide primers identified as SEQ ID No. 1 and SEQ ID No. 2, SEQ ID No. 4 and SEQ ID No. 5, SEQ ID No. 7 and SEQ ID No. 8, and SEQ ID No. 10 and SEQ ID No. 11, and probes with oligonucleotide sequences identified as SEQ ID No. 3, SEQ ID No. 6, SEQ ID No. 9 and SEQ ID No. 12; providing a thermostable DNA polymerase and magnesium salt to the reaction mixture; amplifying the reaction mixture by a PCR reaction; and determining the presence or absence, and quantification of the pathogens by using a fluorescent signal or fluorescence emission specific for each pathogen.

The disclosure relates to an improved method for detecting, locating and monitoring fluid seepage and leakage from a hydraulic work with superior sensitivity. The method includes using a DNA sequenceas the probe to trace the fluid seepage and leakage from a hydraulic work. The probe can be captured and then amplified more than a millionfold by an enzymatic method such as the polymerase chain reaction (PCR) to give a detection signal. Even a single molecule of the DNA probe can be detected by an enzymatic amplification, thus to give superior sensitivity. The improved detection method is applicable to detecting, locating and monitoring fluid seepage and leakage from hydraulic works, the improved method can also be used, for example, to trace the groundwater flow, underground water flow andother liquid flow. Other related methods are also described.

The invention discloses a protein glycosylation detection kit, a detection method and its application, and uses glycosyl tag nucleic acid sequences to modify azide glycosyl groups on the surface of proteins, introduce protein probe nucleic acid sequences, introduce hybridization chain reaction reagents and other technologies Means to achieve specific protein targeting, HCR-based non-enzyme amplification strategy, use it in living cells, specific protein glycosylation detection on cell membrane surface in vivo, imaging protein glycosylation detection kits, detection methods, and in vivo Detection applications in various applications such as in vitro and in vivo.

The invention relates to a novel method for the enzymatic marking of nucleic acid chains (target sequences) by means of nucleotide conjugates. Said nucleotide conjugates are able, under reaction conditions, to specifically bind to the target sequence and integrate into the complementary growing strand by means of a polymerase. The nucleic acid chains marked by such conjugates can be bound to the solid phase. The marking can be carried out parallel to the enzymatic amplification of target sequences.

The invention relates to a novel method for enzymatically marking nucleic acid chains (target sequences) by using nucleotide conjugates. Said nucleotide conjugates are capable of binding specifically to the target sequence under reaction conditions and of being incorporated in the complementary growing strand by means of a polymerase. The nucleic acid chains marked with such conjugates can be bound to the solid phase. The marking can be carried out in parallel with the enzymatic amplification of target sequences.

The invention provides a PCR-SBT method for genotyping human platelet alloantigen system, the method comprising the following steps: preparing human genome DNA; amplifying the gene sequence of the antigen system in human genome DNA; performing double-enzyme synthesis on the amplified product The purified product was subjected to sequencing PCR reaction; the sequencing product was purified by sodium acetate-ethanol precipitation method, followed by capillary electrophoresis sequencing; the obtained sequence was analyzed by software to determine its genotype. The present invention also provides reagents used in the above typing method. The present invention can be used as an independent and widely used identification method, solves the problem of accurate typing of 28 antigen systems of HPA, utilizes the characteristics of high-throughput and accurate results of PCR-SBT for HPA gene typing operation, and can be used in clinical blood transfusion medicine research The related applications in fields such as genetics and genetics will be highly valued, and it has important practical significance for medical research units, pharmaceutical research and reagent development units.

The invention relates to the technical field of nucleic acid detection, in particular to a microRNA detection method based on non-enzymatic amplification and electrochemiluminescence principles. The detection method comprises the steps of: 1) designing DNA hairpin probes, i.e. designing the hairpin H1 and hairpin H2 sequences required by a non-enzymatic amplification system according to a non-enzymatic amplification principle and a detected microRNA sequence; 2) conducting electrochemiluminescence group marking; 3) performing hairpin probe pretreatment; 4) constructing a non-enzymatic amplification detection system; 5) carrying out product cleaning and separation; and 6) conducting signal detection. The method has the characteristics of high sensitivity, accuracy, fastness and simple operation, and the whole amplification process does not involve enzyme, so that the detection cost is greatly reduced.

A technique that uses nanotechnology to electrically detect and identify RNA sequences without the need for using enzymatic amplification methods or fluorescent labels. The technique may be scaled into large multiplexed arrays for high-throughput and rapid screening. The technique is further able to differentiate closely related variants of a given bacterial or viral species or strain. This technique addresses the need for a quick, efficient, and inexpensive bacterial and viral detection and identification system.

A micro-electromechanical platform and array system and methods for identifying microbial species with single molecule electrical conductance measurements are provided. The electromechanical platform has a two-tier actuation mechanism with a long stroke provided by a comb drive and a fine stroke provided by an in-plane flexural actuator. The platform is capable of making contact with a single-molecule, applying a bias, measuring the current, and performing a large number of measurements for statistical analysis. The system is capable of detecting any microbial species without requiring enzymatic amplification by detecting specific RNA sequences, for example. With oligonucleotide target molecules, the conductance is extremely sensitive to the sequence so even single-nucleotide polymorphisms can be identified. The system can also discern between subspecies using the same DNA probe. The system provides reliable, efficient, and inexpensive detection and species-level identification of microorganisms in complex detecting environments.

Provided are methods for validating the presence and character of genomic mutations, particularly single nucleotide polymorphisms (SNPs), by parallel amplification of a portion or the whole genome with at least two different DNA polymerases.

Provided herein is a method for the selective amplification of a target nucleotide sequence located within a nucleic acid molecule, the method comprising contacting the nucleic acid molecule (“template” molecule) with (i) at least one facilitator oligonucleotide, wherein the facilitator oligonucleotide includes at least one modification at or near its 3′ terminus such that 3′ extension from the facilitator oligonucleotide is blocked, and (ii) two or more oligonucleotide primers, at least one of which is an initiator primer modified such that the presence of the modification prematurely terminates complementary strand synthesis, wherein the facilitator oligonucleotide and the initiator primer bind to substantially the same or adjacent regions of the template nucleic acid molecule and the facilitator oligonucleotide further comprises sequences complementary to the target sequence 3′ to the binding location of the initiator primer; and carrying out thermocyclic, enzymatic amplification such that the specific target sequence is selectively amplified.

The present invention discloses the thermodynamic design and concentrations required to design probe compositions with the desired optimal specificity that enables the major allele-containing species (wild-type) to be detected in large stoichiometric excess Enrich, detect, quantify, purify, image and amplify material containing rare alleles of nucleic acids (prevalence < 1%). As an enzyme-free and uniform nucleic acid enrichment composition, this technique is broadly compatible with virtually all nucleic acid-based biotechnologies, including plate readers and fluorometer readouts of nucleic acids, microarrays, PCR and other enzymatic amplification reactions, Fluorescent barcoding, nanoparticle-based purification and quantification, and in situ hybridization imaging.