60 results about "Single nucleotide mutation" patented technology

The invention discloses a fast real-time DNA amplification instrument and a gene mutation detection method. The fast real-time DNA amplification instrument includes a reagent tank and a thermal cycle system. The reagent tank has at least one socket for placing a reaction tube. The thermal circulation system includes a heating body and a radiator, which are distributed on both sides of the reagent tank, and the heating body and the radiator can be driven by the driving mechanism to approach or move away from the reagent tank alternately. The rapid real-time DNA amplification instrument of the present invention realizes the rapid heating and cooling of the reagent by controlling the distance from the heating element and radiator to the reagent tank. This design is not only simple in structure, but also can help the reagent reach the set temperature point faster, Fast thermal cycle is realized, and at the same time, the gene mutation detection method of the present invention only takes 50 minutes from sampling to successful identification of single nucleotide mutations, while similar instruments on the market generally take 2-3 hours, which greatly improves the detection efficiency of gene mutations.

It is intended to provide a probe for detecting a mutation in JAK2 gene capable of detecting a sequence to be detected having a single nucleotide mutation even when the sequence to be detected having such a mutation and a sequence not to be detected having no mutation are present together. At least one oligonucleotide which comprises the same sequence as that of a region up to the 17th to the 22nd base from a first base toward the 5' direction when a cytosine base (C) at position 84 in exon 12 of JAK2 gene composed of a nucleotide sequence represented by SEQ ID NO: 1 is determined to be the first base, and has the cytosine base (C) as the 3' end is used as the probe. By using such a probe in, for example, Tm analysis, even if a sample contains JAK2 gene having a mutation and JAK2 gene having no mutation, the former mutation can be detected. The probe is preferably labeled with a fluorescent dye.

The invention discloses a specific primer for detecting a key gene NtCPS2 single nucleotide polymorphism for tobacco abienol synthesis and a detection method. The specific primer is characterized by comprising two upstream primers CPS2-1F and CPS2-2F and a downstream primer CPS2-R. The detection method comprises the following steps: based on the allele specific PCR (AS-PCR) principle, designing two complementary upstream primers at an SNP mutation site of the key gene NtCPS2 for abienol synthesis, and designing one shared downstream primer at the downstream of the mutation site; and by taking DNA of a tobacco variety to be detected as a template, performing PCR amplification by using two pairs of specific primers, performing electrophoresis detection on the PCR amplified product, and judging whether the PCR amplified product has a characteristic strip of 297bp. The primer pair disclosed by the invention has the advantages of simplicity and convenience in operation, low expense, reliable result, rapid detection speed, distinguishable heterozygosis or homozygosis, and the like, the specific fragrance property breeding process of abienol can be greatly accelerated, the breeding period can be shortened, and the breeding efficiency can be improved.

A method for reducing the efficiency of primer extension by polymerase enzymes when the 3′ end of a primer or growing nucleic acid chain does not hybridize perfectly with the target, for increasing the selectivity of single nucleotide mutation or gene analyses, for suppressing false positive results and for enhancing the fidelity of the amplification of nucleic acid fragments by avoiding the incorporation of mispairs, the method comprising the steps of: (a) obtaining a nucleic acid sample; (b) hybridizing said nucleic acid sample to a primer; (c) subjecting said nucleic acid sample hybridized to a extension reaction by extending a primer with a polymerizing enzyme; and (d) detecting the presence of extension products; wherein the reaction extension mixture medium contains an intercalating agent such as ethidium bromide, dihydroethidium, ethidium homodimer-1, ethidium homodimer-2, acridine, propidium iodide, YOYO®-1 or TOTO®-1. When the intercalating agent is ethidium bromide the concentration is about 1 to 7 g / ml.

A screening method for identifying compounds that alter the fidelity with which the initiation codon in mRNAs is recognized by the translational apparatus in eukaryotes is disclosed. This screening method was used to identify compounds having such activity. Methods of altering the fidelity of initiation codon selection are also disclosed. Methods of treating disorders characterized by single nucleotide mutations in initiation codons using compounds identified by the screening method, as well as methods of treating fungal and parasitic infections and hyperproliferative disorders using compounds identified by the screening method are also disclosed.