12results about How to "Stable amplification" patented technology

The invention discloses a molecular marker for high-throughput identification of a wheat scab site Qfhb7EL and application of the molecular marker. The nucleotide sequences of primers of the molecular marker are as shown in SEQ ID No. 1 to SEQ ID No. 4. Specific primers are designed according to the homologous sequence of the Fhb7 gene in the triticum elongatum, the specific primers and the Actin gene of wheat form a primer pair, population typing is achieved under recognition of different fluorescence, and the reliability of the marker is verified through molecular experiments. Results show that the identification of the wheat scab-resistant site Qfhb7EL in the wheat background can be quickly and accurately realized in a high-throughput manner, and the method has a very high application value for auxiliary selection of new wheat scab-resistant varieties.

The present invention discloses molecular markers related to wheat spike length and number of grains per spike and their applications, belonging to the technical field of crop selection and breeding. The molecular marker is dCAPS-MTA180-DdeI, which marks the SNP site AX-110049563 significantly related to wheat spike length and number of grains per spike, and the nucleotide sequence is shown in SEQ ID NO: 5, and the corresponding genotype of the AX-110049563 site is TT, or the nucleotide sequence is shown in SEQ ID NO: 3 and SEQ ID NO: 4, and the corresponding genotype of the AX-110049563 site is CC. The molecular marker can be widely used in molecular marker-assisted breeding and whole genome selection breeding of wheat spike length and number of grains per spike to accelerate the breeding process.

The present invention discloses a cerebrospinal fluid tumor organoid culture method, which uses primary tumor cells in cerebrospinal fluid for in vitro tumor organoid culture, mixes the primary tumor cells with Matrigel matrix glue, incubates the mixture to solidify the matrix glue, then adds cerebrospinal fluid tumor organoid culture medium for culture, replaces fresh culture medium every two days, and can induce differentiation to form cerebrospinal fluid tumor organoids after 5 days of culture.

The invention relates to the technical field of analog integrated circuits, in particular to a high-precision temperature sensor temperature-sensing front-end circuit based on a chopping technology, and aims to solve the problems of low-frequency 1 / f noise, operational amplifier input offset voltage drift, band-gap reference temperature drift, limited output driving capability and the like existing at the front end of a traditional temperature sensor. The front-end circuit comprises a chopping band-gap reference source circuit, a chopping buffer circuit and a two-phase non-overlapping clock generation circuit, wherein the chopping band-gap reference source circuit adopts a parasitic bipolar PNP transistor with an emitter junction area ratio of 8: 1 as a temperature sensing device, and a high-gain chopping folded cascode operational amplifier is embedded in the chopping band-gap reference source circuit; and stable positive temperature coefficient voltage and low-temperature drift reference voltage are generated. Noise and maladjustment are inhibited from the system architecture level, the temperature measurement precision can reach the sub-degree centigrade level, the loading capacity is high, the method is achieved based on the standard CMOS technology, a special technology and complex temperature calibration are not needed, and the method has the advantages of being low in cost, high in stability and large-scale integration.

This invention belongs to the field of ecological restoration and aquatic plant germplasm identification technology. It provides specific primers and identification methods for *Myriophyllum sieboldii* and *Myriophyllum sieboldii*, based on microsatellite marker (SSR) technology. Ten pairs of highly polymorphic microsatellite primers were screened and used to form five sets of dual PCR primer combinations for amplifying DNA fragments of the target species. Using confirmed *Myriophyllum sieboldii* and *Myriophyllum sieboldii* as basic reference samples, a genetic clustering model was constructed. DNA extraction, PCR amplification, electrophoresis analysis, and UPGMA clustering analysis were performed on the individuals to be identified, thereby achieving accurate species identification of suspected individuals. This invention enables rapid, accurate, and low-cost germplasm identification during the plant's vegetative growth stage and is applicable to wetland ecological restoration, germplasm resource protection, and aquatic vegetation surveys.

The present invention provides a highly polymorphic microsatellite molecular marker of Penaeus vannamei and an application thereof, belonging to the technical field of molecular markers. The present invention is based on 10 microsatellite sequences screened from the genome of Penaeus vannamei, which are numbered Lv‑449, Lv‑460, Lv‑499, Lv‑511, Lv‑442, Lv‑451, Lv‑496, Lv‑491, Lv‑439 and Lv‑482, respectively, and their sequences are shown in SEQ ID No.1 to SEQ ID No.10. The present invention designs microsatellite marker primers for the 10 microsatellite sequences respectively, and the microsatellite marker primers have strong specificity, stable amplification and high polymorphism, and can be used for the genetic diversity and genetic structure analysis of Penaeus vannamei populations and the identification of germplasm resources, providing a powerful tool for the genetic breeding of Penaeus vannamei.

The invention discloses a recessive hereditary powdery mildew resistance gene for watermelon Clpm‑2WF Discovery and application in the breeding of new disease-resistant watermelon varieties. The present invention constructed 6 generations of genetic populations (P1, P2, F1, BC1P1, BC1P2, F2) using the powdery mildew-resistant watermelon inbred line R23 (P1) and the powdery mildew-susceptible inbred line R12 (P2). The powdery mildew resistance was determined to be recessive by inoculating the powdery mildew physiological race 2WF and performing disease identification. Using the extremely resistant and susceptible F2 individual plant resequencing data, the initial positioning interval of the disease-resistant gene was obtained through GWAS analysis, and then the F2:3 family was used for fine positioning. Disease-resistant genes were further screened based on the transcriptome data analysis before and after inoculation of the resistant and susceptible parents. Combining the results of fine positioning and transcriptome analysis, a recessive watermelon powdery mildew resistance gene was identified. Clpm‑2WF Based on the difference sites between resistant and susceptible materials, molecular marker primers were developed for assisted selection of powdery mildew resistant materials, providing genetic materials and methods for breeding powdery mildew resistant varieties of watermelon.

The invention belongs to the technical field of fungus genetic breeding, and particularly relates to a molecular marker associated with the number of shiitake mushrooms and application. The molecular marker is located at the 3865993th site of a chromosome 7 of a mushroom genome and is a section of insertion mutation of 15bp. By utilizing the molecular marker and the detection method provided by the invention, the quantity (mushroom number) of mushroom sporocarp in the fruiting stage can be efficiently judged in the mushroom hypha stage, so that the breeding process is accelerated, and the breeding efficiency is improved. The molecular marker is a universal molecular marker, is slightly limited by genetic materials and is wide in application.

The present application relates to a kind of SSR molecular marker primer set of Mahonia, and its application, the forward primer sequence of the SSR molecular marker primer set includes the sequence as shown in any one of SEQ ID NO:1-7 or as shown in any one of SEQ ID NO:1-7;The reverse primer sequence of the SSR molecular marker primer set includes the sequence as shown in any one of SEQ ID NO:8-14 or as shown in any one of SEQ ID NO:8-14, the SSR molecular marker primer set of the present application is accurately identified long column Mahonia, and it guarantees the authenticity of base original variety for its relevant quality standard research.

The invention belongs to the field of crop breeding science, and discloses a molecular marker of a wheat gibberellic disease resistant gene TaWRKY57 and application of the molecular marker. The invention discloses a CAPS (cleaved amplified polymorphic sequence) marker primer pair of a candidate gene TaWRKY57 of a wheat gibberellic disease resistant gene Qfhi.nau-5B, which comprises WGRB4579-F and WGRB4579-R. After the primer pair is used for carrying out PCR (polymerase chain reaction) amplification and enzyme digestion on wheat variety genome DNA (deoxyribonucleic acid), if three specific fragments with the sizes of 126bp, 135bp and 182bp are obtained, the material is proved to carry the gibberellic disease resistant gene TaWRKY57. The marker has the characteristics of rapidness, accuracy and high efficiency, can be used for determining whether Qfhi.nau-5B exists or not and determining the existence state of Qfhi.nau-5B and predicting the gibberellic disease resistance of wheat, and provides a molecular marker auxiliary means for rapidly screening plants carrying Qfhi.nau-5B and breeding disease-resistant varieties.

The invention discloses a primer set of a KASP molecular marker tightly linked to a major effect QTL of wheat single ear grain weight and its application, and relates to the technical field of molecular marker-assisted breeding. The molecular marker is KASP-AX-109431092, and its primer set comprises 3 primers, and its nucleotide sequences are shown in SEQ ID NO.10, SEQ ID NO.11 and SEQ ID NO.12 respectively. The molecular marker provided by the invention can assist in the screening of high-ear grain weight wheat in wheat breeding, can save breeding costs, improve breeding efficiency, and is of great significance for high-yield wheat breeding.