12 results about "Dna amplification" patented technology

Disclosed herein are DNA amplification methods for quantifying DNA fragments of a target DNA in a sample by size. This can be used, for example, to detect tumor-derived viral DNA in blood sample and distinguish it from larger viral DNA from non-tumor sources. In particular, disclosed herein are methods of detecting, monitoring or treating a human papilloma virus (HPV)-associated malignancy in a subject that involves detecting a presence or absence of at least one circulating tumor-derived HPV DNA in a sample from the subject. Kits for accomplishing the same are also provided.

The application provides a method and application for improving DNA yield in vitro amplification, and belongs to the technical field of biology. The application provides a method for improving DNA yield in vitro amplification, which comprises the following steps: (1) reacting a reaction system at 27-38 DEG C for 40-60 min; (2) adding the reaction liquid into a dialysis bag, adding dialysis liquid into an outer tube, and dialyzing at 30-38 DEG C; (3) supplementing tRNA into the reaction liquid for sealing after dialyzing for 1-3 h; the reaction system in step (1) comprises 5x synthesis enzyme mix, 5x synthesis reaction buffer, NTP, dNTP, a template and water; and the concentration of the dNTP is 0.1-0.5 mM. The method can significantly improve the DNA yield in vitro amplification, the concentration of the amplification product is increased by more than about 134 %, and the core problem of insufficient yield in the in vitro DNA amplification is effectively solved.

This invention discloses a kit for amplifying the whole genome of bovine embryonic cells and its applications. The invention provides a primer set for amplifying the whole genome of bovine embryonic cells, consisting of 20 primers, each 6 nt in length. This invention can utilize 4-8 bovine embryonic cells or genomic DNA down to the pg level to amplify and generate whole-genome DNA amplification products with high coverage and low mismatch rate. These products can be directly adapted to downstream applications such as next-generation sequencing and targeted capture sequencing, while reducing detection costs. Furthermore, the three-step operation process of this invention (cell lysis-lysis termination-isothermal amplification) can complete efficient amplification within 3.5 hours.

A rapid detection and visualization method for triple RPA-LFA targeting necrophilic flies includes the following steps: Step 1: DNA amplification reaction; DNA is extracted from *Gynostemma pentaphyllum*, *Gynostemma spp.*, *Variegata spp.*, *Variegata spp.*, *Bulbus rubrothes*, *Gynostemma pentaphyllum*, *Flyflies fasciatus*, and *Heterodon spp.* using the SteadyPure Universal Genomic DNA Extraction Kit; the concentration and purity of the genomic DNA of the above species are assessed using a Nano Drop 2000 spectrophotometer; the reaction is performed according to the instructions of the DNA isothermal rapid amplification kit; the reaction system uses Buffer A, a primer mixture consisting of 3 pairs of primers; the mixture is incubated in a metal bath to perform the RPA reaction and obtain the amplified product; Step 2: Visual detection; Mixed amplification product: the amplified product is mixed with dd water to obtain a mixed amplification product; the mixed amplification product is dropped onto the LFD test strip sample pad, and then the result is displayed. This invention features no dependence on a sequencer, simple operation, rapid detection, and visualization.

The application belongs to the technical field of biology and relates to a phi29 DNA polymerase mutant with reduced polymerization rate and a preparation method and application thereof. The application designs and provides a mutant of phi29 DNA polymerase. The amino acid residues at at least one of positions 190, 324, 411, 49, 83, 375, 526, 85, 149, 284, 287, 35, 115, 178, 102, 246, 336, 506, 257, 502, 55, 113, 187, 289, 306, 517, 527, 140, 203, 368, 372, 89, 54, 277, 327, 163, 224 in the amino acid sequence of the phi29 DNA polymerase are modified, a new protein with reduced polymerization rate of the polymerase and good polymerase activity is generated, problems caused by the too fast polymerization rate of the phi29 DNA polymerase in practical application are successfully solved, the accuracy, stability and reliability of the phi29 DNA polymerase in DNA amplification, sequencing and other molecular biology experiments are improved, and the sequencing quality is greatly improved.

The application discloses a kind of extraction determination methods of soil nematode digestive tract microorganism, belong to the field of microbial detection technology. Including the following steps: (1) separation extraction nematode: shallow dish method is used to separate nematode in soil, in turn through washing and screening, shallow dish standing, starvation treatment, concentration and preservation operation;(2) digestive tract microorganism extraction: after species identification to nematode, select the nematode individual of specific nutrition group, after surface microorganism elution treatment, extract digestive tract microorganism DNA;(3) DNA amplification: using primer to the V3-V4 region of digestive tract bacteria 16S rRNA gene and the ITS region of fungal ITS gene is carried out PCR amplification;(4) sequencing and data analysis: after purification and quantification to PCR amplification product, using Illumina MiSeq platform carries out double-end sequencing, original data is split, quality filtering, sequence splicing, chimeric sequence removal, ASV clustering and species annotation processing.

The application discloses a reagent, a method and application for improving PCR inhibitor tolerance, and belongs to the technical field of molecular biology. The application combines heat-sensitive uracil DNA glycosylase and deoxyuridine triphosphate, and applies the combination as a PCR anti-inhibitor in DNA amplification of a whole blood sample. Experiments prove that the combination can improve the tolerance of a PCR reaction system to whole blood inhibitors, reduce the false negative rate, and improve the accuracy and reliability of detection. Meanwhile, the method for performing PCR reaction by using the combination is simple in operation, low in cost, and high in nucleic acid detection sensitivity. Therefore, the reagent and the method have good application prospects in the detection of nucleic acids extracted from a whole blood sample.

This invention belongs to the field of bioengineering technology, specifically relating to a recombinant plasmid for tylosin synthesis, an engineered bacterial strain, and its preparation method. The genetically engineered bacterial strain (Streptomyces freundii G3-F2) was deposited at the China Center for Type Culture Collection (CCTCC) on February 24, 2025, with accession number CCTCC NO: M 2025278. This invention achieves site-specific recombination, utilizes an optimized ZouA gene DNA amplification system to construct the recombinant plasmid, and realizes… tulle Multiple copies of the F gene were increased, resulting in the preparation of *Streptomyces freundii* G3-F2. This significantly improved the rate of methyltransferase enzymatic reaction and achieved efficient conversion of component C. Combined with low-temperature fermentation, the rate and efficiency of tylosin synthesis were greatly enhanced. The strain achieved a titer of 18500 U / mL in a 50L tank under 28℃ low-temperature culture conditions, which can significantly reduce production costs.

This invention discloses a primer combination for rapid detection of *Beetroot Cystus* using convective PCR fluorescence, and a kit for rapid detection of *Beetroot Cystus* using convective PCR fluorescence comprising the primer combination. This invention designs upstream and downstream primers and fluorescent probes targeting a specific gene fragment of the *Beetroot Cystus* mitochondrial COI gene, and provides a detection kit containing specific primers, probes, and reaction solutions. This enables rapid amplification and fluorescence detection of the target region of *Beetroot Cystus*. Utilizing a rapid nucleic acid extraction reagent, detection can be completed within 30 minutes, greatly improving the efficiency of DNA amplification and fluorescence detection. The detection is highly accurate, sensitive, simple to operate, time-saving, reproducible, and practical.

This invention pertains in general to probes for detecting and / or quantifying double- stranded DNA. The invention particularly relates to luminescence based probes, and methods of using such probes to detect or quantify double-stranded DNA, and in particular their use in real time DNA amplification and quantification reactions. The invention further describes compositions including the probes suitable for the described uses and methods.

The present invention belongs to the field of biotechnology, and relates to a phi29 DNA polymerase mutant having a reduced polymerization rate, a preparation method therefore, and a use thereof. The present invention designs mutants of phi29 DNA polymerase, and by means of modifying amino acid residues at at least one of positions 190, 324, 411, 49, 83, 375, 526, 85, 149, 284, 287, 35, 115, 178, 102, 246, 336, 506, 257, 502, 55, 113, 187, 289, 306, 517, 527, 140, 203, 368, 372, 89, 54, 277, 327, 163, and 224 in the amino acid sequence of phi29 DNA polymerase, generates novel proteins that reduce the polymerization rate of the polymerase while maintaining good polymerase activity, successfully solving problems caused by the excessively fast polymerization rate of phi29 DNA polymerase in practical applications, improving the accuracy, stability, and reliability thereof in DNA amplification, sequencing, and other molecular biology experiments, and greatly enhancing sequencing quality.

This invention pertains in general to probes for detecting and / or quantifying double-stranded DNA. The invention particularly relates to luminescence based probes, and methods of using such probes to detect or quantify double-stranded DNA, and in particular their use in real time DNA amplification and quantification reactions. The invention further describes compositions including the probes suitable for the described uses and methods.