93results about How to "Amplification realized" patented technology

The invention belongs to the technical field of PCR (polymerase chain reaction), and particularly relates to a digital PCR chip with silicon substrate arrays and micro-reaction pools and a method for manufacturing the digital PCR chip. The digital PCR chip mainly comprises an upper cover and a chip. Micro-pores which are arrayed to form honeycombs are formed in the chip, and the upper cover is fixed onto a chip groove. The method includes particular steps of selecting a silicon chip with a single polished surface; cleaning the silicon chip; coating a layer of uniform photoresist on the polished surface of the silicon chip in a spinning manner; forming circular graphic arrays by means of exposure; etching silicon by the aid of a dry process in masks, namely, the circular graphic arrays to form micro-pore structures; removing the photoresist and scribing the chip to completely manufacture the digital PCR chip.

The invention belongs to the technology field of the cellular immunology, and particularly relates to a method for the dominant amplification of in vitro peripheral blood nature killer cells with a large amount. The method comprises the steps as follows: (1) separating single nucleus cell from human or animal peripheral blood; (2) suspending the single nucleus cell in an RPMI 1640 culture solution containing 5%-15% autologous serum and pretreating with methyl-Beta-cyclodextrin with an effective concentration of 1-4mmol/L for 36-60 hours; and (3) adding recombined interleukin 2, and amplification-culturing in an RPMI 1640 culture solution containing 5% of new-born calf serum and 5% of autologous serum for above 10 days. The method has the advantages that the blood sample amount is small; the cost is low; the operation is convenient; and the amplification multiple is high, and the nature killer cells can be amplified by 392-1752 times in a short term.

The invention discloses a loop-mediated isothermal amplification primer composition capable of detecting colletotrichum truncatum and application thereof. The primer composition comprises a forward outer primer Rpb1-F3 as shown in SEQ ID NO.2, a reverse outer primer Rpb1-B3 as shown in SEQ ID NO.3, a forward inner primer Rpb1-FIP as shown in SEQ ID NO.4, a reverse inner primer Rpb1-BIP as shown in SEQ ID NO.5 and a loop primer Rpb1-LF as shown in SEQ ID NO.6. By the primer composition, the problems that the biological detection method requires long period, wastes time and energy and is tedious and poor in specificity and a thermal cycling instrument is needed and colletotrichum truncatum cannot be rapidly detected by virtue of the PCR detection technology in the prior art are solved and the primer composition has the advantages of short detection period (only 70 minutes), strong specificity and high sensitivity, and the detection results can be visually observed.

The invention relates to a nucleic acid detection kit for RNA isothermal amplification avian influenza virus H7N9 (2013). The kit includes a virus preservation solution, a nucleic acid extraction solution, a washing liquid, an H7N9 reaction liquid (2013), an H7 detection liquid, an N9 detection liquid, an SAT enzyme liquid, an H7N9 (2013) positive control reagent, and an H7N9 (2013) negative control reagent. The detection kit provided by the invention has the characteristics of high specificity, high sensitivity, low pollution and rapid detection, and plays an important role in clinical diagnosis of early infection of avian influenza virus H7N9 (2013).

The present invention is one optimized long segment nucleic acid PCR amplification system based on nanometer metal particle, and belongs to the field of molecular biology technology. One Long-PCR system is configured on the basis of available PCR amplification process, and the Long-PCR system has at least 10 vol% of 0.01 wt% concentration nanometer metal colloid solution or nanometer metal particles in the ultimate concentration not lower than 0.001 wt% added into the PCR system. The present invention has simple operation, obvious optimizing effect, low cost and wide application.

The invention discloses an RDA method and kit for rapidly detecting respiratory syncytial viruses. The kit comprise a specific primer pair and an RDA fluorescence labeling probe to realize safe, specific, sensitive, simple and convenient detection of the respiratory syncytial viruses (RSV) and overcome the defects of the existing traditional detection technology. According to the kit provided by the invention, a nucleic acid extraction step can be omitted, the respiratory syncytial viruses can be detected at the constant temperature of 37-42 DEG C within 20 minutes, the specificity is 100%, and the kit is very suitable for on-site rapid detection. Compared with a common PCR method, the RDA fluorescence method has the advantages that the reaction is performed at constant temperature, the temperature change is not needed, complex instruments are not needed, and the reaction time is short. Therefore, the method and the kit have the characteristics of simplicity and quickness in operation,good specificity, high sensitivity, low cost and the like, provide an effective technical means for on-site quick detection and screening of the respiratory syncytial viruses, and have a wide application prospect.

The invention discloses a loop-mediated isothermal amplification (LAMP) detection method for detecting strawberry anthracnose in the seedling stage and an LAMP primer used in the method; and the method can be used for quickly detecting whether strawberry seedlings carry germs or not on the site. The invention also discloses an LAMP detection method for detecting anti-QoI germicide G143A genotype strawberry colletotrichum gloeosporioides and an LAMP primer used in the method; and the method can be used for specifically detecting strawberry colletotrichum gloeosporioides with high resistance toQoI germicides. The method disclosed by the invention has the characteristics of simplicity, quickness, low cost, high sensitivity and the like, can greatly improve the detection efficiency, and has important practical significance for seedling stage detection and drug resistance epidemic early warning of strawberry anthracnose.

The invention discloses a method for molecular detection of Phytophthora cambivora(Petri)Buisman with the loop-mediated isothermal amplification (LAMP) technique based on color differentiation, and primers. The sequences of the primers are shown from SEQ ID NO.1 to SEQ ID NO.6. According to the detection system, under the isothermal condition of 62 DEG C, Phytophthora cambivora(Petri)Buisman can be detected quickly, conveniently and efficiently with high specificity and sensitivity, no complicated instrument is needed, a new technical platform is provided for detection of Phytophthora cambivora(Petri)Buisman, and the requirement for field detection of Phytophthora cambivora(Petri)Buisman can be well met. The detection system is suitable for inspection and quarantine of entry/exit plants and plant products, and investigation, rapid diagnosis and monitoring of diseases, and is of great importance in preventing Phytophthora cambivora(Petri)Buisman from entering China. Meanwhile, the establishment of the system provides technical guidance and theoretical basis for detection of other pathogenic bacteria.

The invention discloses a carbon fiber tow layered conveying device which comprises a tow collecting device and a tow expanding device. The tow collecting device is provided with a machine frame, a first rotating guide roller and a second transmission guide roller are arranged on the upper portion of one side of the machine frame, a first guide roller is fixed below a second rotating guide roller on the machine frame, a second guide roller is fixed below the first guide roller on the machine frame, a first supporting roller and a second supporting roller are fixed on the upper portion of the other side of the machine frame, a third guide roller is fixed below the first supporting roller on the machine frame, a fourth guide roller is fixed below the third guide roller on the machine frame, and the tow collecting device and the tow expanding device are of the same symmetrical structure. The tow collecting device and the tow expanding device are respectively placed on the front of a carbide furnace and on the back of the carbide furnace, the tow collecting device is capable of achieving layering and collecting of tows, and the tow expanding device is capable of achieving expanding of the tows.

The invention discloses an RDA method and a kit for rapidly detecting influenza A virus. The kit comprises a specific primer group and an RDA fluorescence labeling probe so as to realize safe, specific, sensitive, simple and convenient detection of the influenza A virus (FluA), and to make up for the defects of existing traditional detection technologies. According to the kit provided by the invention, a nucleic acid extraction step can be omitted, the influenza A virus can be detected within 20 minutes at a constant temperature of 37-42 DEG C, the specificity is 100%, and the kit is very suitable for on-site rapid detection. Compared with common PCR methods, the RDA fluorescence method has the advantages that: the reaction is carried out at constant temperature, the temperature change isnot needed, complex instruments are not needed, and the reaction time is short. Therefore, the method and the kit provided by the invention have the characteristics of simple and rapid operation, goodspecificity, high sensitivity, low cost and the like, provide an effective a technical means for on-site rapid detection and screening of the influenza A virus, and have a wide application prospect.

To achieve the above object, the present invention discloses a fusion Taq DNA polymerase, which is formed by fusing a DNA single-strand binding protein Sac7d or a DNA single-strand binding protein sso7d with a wild-type Taq DNA polymerase, respectively. The Taq DNA polymerase provided by the invention enhances the affinity with the template chain in the amplification process so as to improve the amplification efficiency, realizes the amplification of difficult templates, has stable enzyme activity and can be directly amplified by using blood as a template.

The invention discloses an RDA method and a kit for rapidly detecting feline panleukopenia virus (FPV). The kit for rapidly detecting the FPV comprises a specific primer pair and an RDA fluorescent label probe so as to realize safe, specific, sensitive, simple and convenient detection of the FPV, thereby making up the defects of the existing traditional detection technology. The kit provided by the invention can save the step of nucleic acid extraction, and is capable of detecting the FPV within 20 min at a constant temperature; and moreover, the kit has a specificity of 100%. Compared with acommon PCR method, the RDA fluorescence method has the advantages of being capable of carrying out reaction at a constant temperature, free of requirements for temperature change and complex instruments, and short in reaction time. Therefore, the kit is suitable for on-site rapid detection. The method and the kit thereof disclosed by the invention have the characteristics of being simple and rapidin operation, excellent in specificity, high in sensitivity, low in cost and the like; and thus, an effective technical means is provided for on-site rapid detection and screening of the FPV. The method and the kit have a wide application prospect.

The invention relates to the field of submerged arc furnace equipment, in particular to an automatic electrode paste adding device and method for submerged arc furnaces. By means of the device, an electrode paste storage bin, a conveying device capable of achieving long-distance conveying and a servo system single beam crane are combined, so that electrode paste can be injected to multiple sets ofsubmerged arc furnaces automatically, the electrode paste injecting efficiency is improved, in addition, the electrode paste stored remotely can be conveyed to the submerged arc furnace, and the field utilization rate is increased. The invention further provides an automatic electrode paste adding method for the submerged arc furnaces, an electric control system and a monitoring device are used for conducting programming control and real-time detection, the electrode paste injecting process is more intelligent, and the work efficiency is improved.

The invention provides a generation method for general amplification of an intelligent automobile recombination scene, which comprises the following steps: determining key elements, including static elements and dynamic elements, for scene generation according to test requirements, and setting related fields for each kind of information; determining a value domain of each element field and whetherthe element fields have correlation or not by using a statistical analysis method based on historical data of a natural driving database, and randomly selecting and traversing elements without correlation in the value domain; for elements with correlation, obtaining constraint conditions through a statistical analysis method, and carrying out selection in a value domain corresponding to the constraint conditions of the fields. According to the generation method for universal amplification of the intelligent automobile recombination scene, the recombination scene can be rapidly generated, scene amplification is achieved, and the purpose of covering all possible working conditions is achieved.

The invention discloses a loop-mediated isothermal amplification primer for detecting botrytis cinerea containing BCbil43/144 intron. The invention further simultaneously discloses a loop-mediated isothermal amplification kit for detecting the botrytis cinerea containing the BCbil43/144 intron. Besides the LAMP (loop-mediated isothermal amplification) primer composition, the kit further comprises 10*ThermoPol Buffer, dNTPs, MgCl2, betaine, hydroxynaphthol blue, 8U/mu L Bst DNA polymerase and ddH2O. By adopting the loop-mediated isothermal amplification primer and the loop-mediated isothermal amplification kit which are disclosed by the invention, rapid detection on the botrytis cinerea containing the BCbil43/144 intron can be implemented in the field, so that scientific medicine application for preventing and treating the botrytis cinerea is timely guided.

The invention provides a multiplex PCR primer group and next-generation sequencing database building method for MLST (multilocus sequence typing) tracing of vibrio parahaemolyticus, and belongs to thetechnical field of microbiological detection. The multiplex PCR primer group for the MLST tracing of the vibrio parahaemolyticus comprises a primer for gyrB gene amplification, a primer for dnaE geneamplification, a primer for dtdS gene amplification, a primer for pyrC gene amplification, a primer for tnaA gene amplification, a primer for pntA gene amplification and one of a primer for recA-1 gene amplification and a primer for recA-2 gene amplification. The next-generation sequencing based database building method is established by multiplex PCR amplification for the seven genes, amplification and sequencing of the two types of RecA genes can be realized, and a sequencing result is relatively consistent with a first-generation sequencing result; and meanwhile, amplification of all genescan be realized by one-time PCR, the method can be directly applied to database building, and the operation is simple.

The invention discloses a Mycoplasma pneumoniae detection method based on PCR (polymerase chain reaction)-gold-magnetic nano immunochromatography assay, which comprises the specific steps of designinga specific primer modified with digoxin and biotin according to 16S rRNA nucleotide sequence of Mycoplasma pneumoniae; extracting DNA of a sample under test, and performing PCR amplification by usingthe DNA of the sample under test as a template and the specific primer as a primer to obtain a MP target fragment amplification product of the sample under test; detecting the amplification product with a Mycoplasma pneumoniae detection gold-magnetic immunochromatographic test strip. The Mycoplasma pneumoniae detection method based on PCR-gold-magnetic nano immunochromatography assay is simple toperform and is suitable for detecting Mycoplasma pneumoniae quickly.