31results about How to "No sequencing required" patented technology

The invention relates to a related specie identification technology, in particular to a method for identifying related species of an oyster. The method specifically comprises the following steps of: by using an oyster genome DNA (Deoxyribonucleic Acid) as a template, designing a primer according to a tag sequence of the oyster specie genome; carrying out PCR (Polymerase Chain Reaction) amplification on the primer; and distinguishing oyster species by using Tm value difference in a high-resolution melting curve of an amplification product. Through the adoption of the method provided by the invention, the species can be identified rapidly, economically, simply and conveniently; and compared with a tag sequence sequencing result, an identifying result of the method provided by the invention is 100% in accuracy.

The invention relates to a group of specific primers and a method for identifying stiff silkworms, belonging to the technical field of identification of traditional Chinese medicines. According to themethod, after a stiff silkworm sample is crushed, the deoxyribonucleic acid (DNA) in domestic silkworms and the DNA in fungi are extracted at the same time by using a CTAB method; the extracted DNA is taken as a template, specific nucleotide sequences, which are designed and screened, are used as primers for performing polymerase chain reaction (PCR) amplification, and the products are then subjected to agarose gel electrophoresis analysis and gel imaging system detection, so that the authenticity and adulteration of the stiff silkworms can be finally detected based on the detection results.The method provided by the invention can be used for judging whether the sample is a stiff silkworm fake product or adulterated product, is simple to operate, does not needing sequencing, is high in specificity, rapid and accurate, and has unique advantages and important application value in the authenticity identification of stiff silkworm samples.

The invention discloses a method for authenticating and distinguishing fusarium culture causing soybean root rot. The method is characterized in that: fusarium solani and other 3 kinds of fusarium areseparated by using 3 percent agarose gel electrophoresis according to morphology and a genetic evolution tree authentication result built by an ITS sequence, and in terms of an amplified ITS sequencelength; comparing the ITS sequences of the rest 3 kinds of fusarium, finding out a restriction enzyme cutting site BspCNI, AvaI and PstI of a specific-limited incision enzyme; forming different CAPSmolecular markers with ITS sequences obtained by performing enzymatic hydrolysis and amplification on different combinations of the 3 kinds of enzymes, separating the molecular markers with 1.5 percent agarose gel electrophoresis in order to separate the 3 kinds of fusarium. By adopting the method, the authentication and distinguishment of the fusarium culture can be realized by means of the agarose gel electrophoresis and the CAPS molecular markers, the method is suitable for large-scale and high-flux culture authentication and distinguishment, in particular to the rapid authentication and distinguishment of the main fusarium causing soybean root rot in the national soybean production areas.

The invention relates to an ITS2 molecular marker identifying method for aquaculture of catfish hybrids. Southern catfishes, catfishes and hybrids of the southern catfishes and the catfishes serve as objects of study, ITS2 gene sections in catfishes are selected to serve as molecular markers, the length difference the IST2 gene sections of the southern catfishes and the catfishes, and the southern catfishes, the catfishes and hybrids of the southern catfishes and the catfishes are identified successfully. The method has the advantages of being fast in experiment, convenient to operate, visual in identification and free from sequencing.

The present invention discloses a kit for detecting one pathogenic mutation site in male infertility Galntl5 gene, and a PCR amplification method thereof. The method is characterized in that mutation detection primers resisting 3'-5'exonuclease digestion modification are introduced into a high fidelity DNA involved PCR reaction so as to improve the PCR specificity, and the product band of the gel electrophoresis can be directly observed so as to determine the genotype of the corresponding mutation site of the Galntl5 gene. According to the present invention, the detection kit has characteristics of low instrument requirements, simple operation, economy, no requirement of sequencing, and capability of being operated in general molecular biology laboratories.

The invention discloses a kit for detecting two pathogenic mutation loci in a TAB2 gene of a human congenital heart disease, and a PCR (Polymerase Chain Reaction) amplification method thereof. The method is characterized in that a mutation detection primer modified by an anti-3'-5' excision enzyme in an enzyme digestion manner is applied into the PCR in which a high-fidelity DNA (Deoxyribose Nucleic Acid) polymerase is participated, so that the PCR specificity is improved; thus, the genetype of the corresponding mutation loci of the TAB2 gene can be judged by directly observing whether a gel electrophoresis product stripe exists or not. The kit has a low requirement to instruments and equipment, is simple to operate and economic and has no need of sequence measuring. As a result, the kit can be operated in an ordinary molecular biology laboratory.

The invention discloses an SNP marker for identifying Corydalis turtschaninovii Bess., an allele-specific PCR process and application, and relates to the technical field of biology. The SNP marker andthe process are provided based on a problem that there is no specific identification method for Corydalis turtschaninovii Bess.. The invention discloses the SNP marker for identifying Corydalis turtschaninovii Bess., the application of the SNP marker, a specific primer pair T1 for detecting Corydalis turtschaninovii Bess. based on allele PCR detection, and the allele-specific PCR process for Corydalis turtschaninovii Bess. detection. Beneficial effects of the SNP marker, the application, the process, and the specific primer pair are that accurate identification of the Corydalis turtschaninovii Bess. and processing products thereof are achieved, and advantages including high accuracy, high stability, high specificity, no need of sequencing, and no influence caused by medicinal parts are achieved.

The present invention discloses a kit for detecting one pathogenic mutation site in male infertility Kif18a gene, and a PCR amplification method thereof. The method is characterized in that mutation detection primers resisting 3'-5'exonuclease digestion modification are introduced into a high fidelity DNA involved PCR reaction so as to improve the PCR specificity, and the product band of the gel electrophoresis can be directly observed so as to determine the genotype of the corresponding mutation site of the Kif18a gene. According to the present invention, the detection kit has characteristics of low instrument requirements, simple operation, economy, no requirement of sequencing, and capability of being operated in general molecular biology laboratories.

The invention relates to a multiple PCR (polymerase chain reaction) kit for simultaneously detecting anopheles peditaeniatus, anopheles lesteri and anopheles barbirostris. The kit comprises specific primers for the anopheles peditaeniatus, the anopheles lesteri and the anopheles barbirostris, 10 * buffer, dNTPs, MgCl2, Taq enzymes, template DNA and sterile double distilled water, wherein the sequences of the specific primers for the anopheles peditaeniatus, the anopheles lesteri and the anopheles barbirostris are respectively shown as SEQ ID NO.5-6, SEQ ID NO.15-16 and SEQ ID NO.33-34. The multiple PCR detection kit disclosed by the invention can provide a qualitative detection result within 3 hours after a DNA sample is received, saves cost and time, has low requirements for personnel andequipment, and is an effective tool for detecting the anopheles peditaeniatus, the anopheles lesteri and the anopheles barbirostris.

A method for quickly detecting the gene site mutation of epidermal growth fact receptor for non-small-cell lung cancer includes such steps as providing DNA, PCR amplifying by specially designed three groups of 4 primers, gel electrophoresis, and abserving result.

The invention provides the primer and method for fast detection of chorea IT15 gene CAG repeat sequence expansion mutation, characterized in that a specially designed primer is employed, and PCR expansion is carried twice to expand DNA amount to gel electrophoresis observable degree, finally gel electrophoresis is utilized to directly observe the situation of IT15 gene CAG repeat sequence dynamic mutation.

The invention relates to a kit for simultaneous detection of Anopheles splendidus, Anopheles maculatus and Anopheles aconitus by multiple PCR. The kit includes specific primers of Anopheles splendidus, Anopheles maculatus and Anopheles aconitus, 10xbuffer, dNTPs, MgCl2, Taq enzyme, template DNA and sterile double distilled water, wherein sequences of the specific primers of Anopheles splendidus, Anopheles maculatus and Anopheles aconitus are shown as SEQ ID NO.5-6, SEQ ID NO. 13-14, and SEQ ID NO 25-26. The multiple PCR detection kit can provide qualitative detection results within 3 hours after receiving DNA samples, the cost and time are saved, the requirements on personnel and equipment are low, and the multiple PCR detection kit is an effective tool for detecting Anopheles splendidus,Anopheles maculatus and Anopheles aconitus.