233results about How to "Good amplification effect" patented technology

The invention discloses a non-small cell lung cancer targeted therapy gene detection method, and belongs to the field of gene detection. The method for detecting 466 mutations of 12 oncogenes is developed by multiplex PCR and high throughput sequencing technologies, wherein the oncogenes are AKT1, ALK, BRAF, EGFR, ERBB4, FGFR1, FGFR2, FGFR3 , KRAS, MET, PIK3CA, and PTEN, and the mutations may be substitutions, insertions and / or deletions of one or more bases. The detection method provided by the invention has the advantages of high detection sensitivity of up to 0.01%, and clear and objective detection results, can be directly used for reflecting specific mutation sites of the relevant gene, directly used for guiding clinical non-small cell lung cancer targeted dosage, and used for early diagnosis or auxiliary diagnosis and screening of cancers as well as post-cancer surveillance.

The invention discloses a primer, a kit and a method for determining a lung cancer gene mutation site based on a high-flux sequencing technology, which belongs to the field of biological molecular detection. The invention discloses a method and a kit for determining a lung cancer gene mutation site based on a high-flux sequencing technology. The method comprises the following steps: extracting tumor tissue DNA; designing a panel of a lung cancer targeting treatment molecular diagnosis associated gene; performing the PCR primer amplification; and establishing a library, and performing the high-flux sequencing. The specific primer and the kit for determining the lung cancer gene mutation site are used for detecting 316 mutation situations of 16 oncogenes, and the mutation can be the replacement, insertion and/or deletion of one or more alkaline groups. The sensitivity of the detection method and the kit of the invention can reach up to 1 percent, the detection result is definite and objective and can directly reflect the specific mutation site of the reaction associated gene and has important significance for early diagnosis or auxiliary diagnosis and screening of the cancer and theprognosis monitoring of the cancer.

The invention discloses a kit for simultaneously detecting nucleic acids of seven respiratory pathogens and application of the kit. The kit is based on a double amplification technology (RNA isothermal amplification and multi-biotin signal amplification), and can detect H1N1/H3N2 type influenza A viruses, influenza B viruses, respiratory syncytial viruses, 1/2/3 type human parainfluenza viruses, B/E type adenoviruses, mycoplasma pneumoniae and chlamydia pneumoniae. The kit of the invention does not need RNA extraction, and is not prone to pollution, high in sensitivity and good in specificityin the process of detection, and can be widely applied to detection of the nucleic acids of the above seven respiratory pathogens.

The invention discloses a specific primer pair based on a rbcL gene and used for identifying land plant species and an application thereof. The specific primer pair provided by the invention is composed of a single stranded DNA A and a single stranded DNA B, wherein the single stranded DNA A is 15-40bp and comprises a DNA fragment of the DNA fragments as shown by a sequence 1 of a sequence table; and the single stranded DNA B is 15-40bp and comprises a DNA fragment of the DNA fragments as shown by a sequence 2 of the sequence table. The single stranded DNA A can be the DNA fragment as shown by the sequence 1 of the sequence table; and the single stranded DNA B can the DNA fragment as shown by the sequence 2 of the sequence table. The specific primer pair provided by the invention can be used for developing a universal kit, effectively identifying the land plant species and prompting the development and social service of plant DNA bar code.

The invention discloses a mixed type hot-start DNA polymerase composition, a PCR amplification kit, and applications thereof. The composition comprises Taq DNA polymerase, an anti-Taq antibody, and mutant type KOD DNA polymerase. The anti-Taq antibody is capable of specifically being combined with Taq DNA polymerase. The 147th site of wild type KOD DNA polymerase is histidine, and the 147th site of mutant type KOD DNA polymerase is lysine. The experiment results show that the mismatch rate is low when the mixed type hot-start DNA polymerase composition is used for PCR amplification, the composition has the dual characteristics of high efficient amplification and high fidelity and is heat resistant in amplification, and the amplification effect is good no matter for a long DNA segment, a template with a low amount, or a template with a high GC content.

The invention discloses nucleic acid and a kit for detecting G1165C polymorphism of the human ADRB1 gene, and also establishes a method for detecting the G1165C polymorphism of the human ADRB1 gene, which is strong in specificity, high in sensitivity, high in degree of accuracy, and easy to operate. The kit for detecting the G1165C polymorphism of the human ADRB1 gene is applicable to the selection for various clinical samples, and has the remarkable advantages of strong specificity, high sensitivity, short experimental period, operation simplicity, safety and non-toxicity, low cost, and the like. The detection method provided by the invention adopts the completely closed tube operation, and is easy, convenient and rapid to operate, the detection result is obtained by directly directing fluorescence signal values during the PCR process, the PCR aftertreatment or electrophoresis detection is not needed, the large possibility of pollution and false positive property during the conventional PCR technology are overcome, the difficulty of non-specific amplification can be effectively avoided, and the detection method is suitable for detection samples in large batches.

The present invention relates to a culture system and method for amplifying hematopoietic stem cells and/or hematopoietic progenitor cells, hematopoietic stem cells, and hematopoietic progenitor cells. The culture system comprises: a basal medium suitable for stem cell expansion; and a JNK signaling pathway inhibitor. The culture system provided by the invention can expand hematopoietic stem cellsin vitro, and the amplification effect thereof is obvious, and the CD34+CD45RA-labeled hematopoietic stem cells can be amplified nearly 100 times.

The invention provides fusion type Taq DNA polymerase as well as a preparation method and application thereof. The fusion type Taq DNA polymerase comprises a Taq DNA polymerase mutant and DNA bindingprotein, wherein the Taq DNA polymerase mutant is protein obtained by carrying out substitution mutation on one or more sites of a helix-hairpin-helix DNA binding region of wild type Taq DNA polymerase. The fusion type Taq DNA polymerase has enhanced nucleic acid binding capacity, amplification speed and impurity tolerance, has good resistance to inhibition of reverse transcriptase, and has good amplification performance for trace templates, and a biological sample can be directly detected without a nucleic acid extraction step; and a constructed one-step RT-qPCR kit is high in detection speed, high in sensitivity and good in accuracy, and has a wide application prospect in the field of RNA virus detection.

The invention discloses a fluorescent constant-temperature amplification technique. In water used as solvent for a fluorescent constant-temperature amplification reagent used in the technique, Tris-buffer solution, magnesium acetate, dithiothreitol, betaine, Tween 20, DMSO, mycose, PEG, BSA, dNTPs, and proper amounts of fluorescent dye, polymerase, single strand DNA-binding protein and recombinase are added. A constant-temperature amplification system of the invention is dependent of high-energy molecules such as ATP and creatine phosphate; compared with existing constant-temperature amplification systems, the constant-temperature amplification system is simpler in composition, lower in cost and more convenient to use. The constant-temperature amplification system is capable of amplifying DNA and RNA sequences, is applicable to amplification of complex templates stably and quickly, capable of finishing an amplification reaction within 30min, has high amplification sensitivity and accuracy without rare occurrence of mispairing, and has good amplification effect. The constant-temperature amplification system is applicable to real-time quantification and has a wider range of application.

The invention discloses a Taq DNA polymerase mutant and application thereof. The Taq DNA polymerase mutant has amino acid substitutions at one or more of the following amino acid positions in the sequence shown as SEQ ID NO. 1; and the various amino acid substitutions are represented in triplets as 'letters-numbers-letters', wherein the numbers reflect position of an amino acid mutation, the letters before the numbers are corresponding to an amino acid involved in the mutation, and the letters after the numbers reflect an amino acids used to replace the amino acid corresponding to the lettersbefore the numbers. The Taq DNA polymerase mutant disclosed by the invention is capable of changing configuration of an enzyme, thereby improving tolerance of the enzyme; and thus, the mutant can be very well applied in blood sample amplification.

The invention provides molecular markerd of a maize chloroplast genome and its application in cultivar identification. The molecular markerd comprise 57 SNP markers and 6 InDel markers. The 63 molecular markers and primers developed according to the markers can be used in any one of the following: (1) constructing a corn variety chloroplast DNA-SNP and InDel fingerprint database, (2) carrying outcorn maternal line traceability analysis, and (3) performing reciprocal cross identification on corn samples. Through the application of the chloroplast polymorphic primer composition, the range of available marker sites of the corn is expanded at the genome level and the new ideas and technical means are provided for the study of maize variety and germplasm resource identification, genetic relationship evaluation and cytoplasmic genetic characteristics.

The invention discloses an RT-PCR-HRM or PCR-HRM primer, reagent and method for fast distinguishing four kinds of serotype dengue viruses. According to the genomic sequence alignment result of the four kinds of serotype dengue viruses, three primers are designed, RNA is adopted as a template, an LC green dye is added for carrying out a one-step RT-PCR, or a built plasmid containing a dengue virus 280bp basic group is adopted as a template and the LC green dye is added for carrying out PCR amplification, the amplification product is subjected to HRM analysis, and the four kinds of serotype dengue viruses can be obviously distinguished through an HRM standard curve and a peak type figure. The method is easy to operate, the reagent is low in cost, high in detecting sensitivity and reliable in parting result, and the method is an innovative method for detecting and parting the four kinds of serotype dengue viruses.

The invention relates to an SSR (Simple Sequence Repeat) core primer group and a method thereof for identifying a tea variety, and belongs to the technical field of biology. The method is characterized by comprising the following steps: (1) material selection; (2) genome DNA extraction; (3) PCR (Polymerase Chain Reaction) amplification of a target product; (4) electrophoretic separation of a PCR product; (5) silver staining development; (6) data analysis; and (7) result judgment. The SSR core primer group and the method thereof for identifying the tea variety, which are disclosed by the invention, have the characteristics of easiness and convenience for operation, accuracy in result, high repeatability, high identification capacity, low cost and the like and are suitable for identifying the tea variety.

The invention discloses a method for identifying donkey skin from counterfeit species by applying mitochondria COI (cytochrome oxidase subunit I) sequence segments and a specific primer for amplifying the segments, belonging to the technical field of identification of Chinese medicine. According to the method disclosed by the invention, COI sequences of the donkey skin and the counterfeit species are amplified by using an autonomously designed primer, the donkey skin is distinguished from pigskin or cowhide by judging whether a PCR (Polymerase Chain Reaction) product exists in an amplified band in gel electrophoresis or not, and mitochondria COI sequences of the donkey skin from multiple sources and common counterfeit species are subjected to sequence assembly and compared, and then the donkey skin and counterfeit species thereof are intuitively identified through a K2P distance method. The method disclosed by the invention is convenient and rapid and high in accuracy and has the capability of well ensuring the quality of donkey-hide gelatin and donkey skin medicinal materials.

The invention discloses a vibration table fixture capable of accurately adjusting the center of gravity, comprising a vibration table top and four subsidiary fixtures in same structures, wherein each subsidiary fixture comprises two guiding supporting plates, two sliding bars, a supporting block, screws, a sliding block, a supporting disc and a base; the four subsidiary fixtures are symmetrically distributed by using a center hole of the vibration table top as the center; the center lines of the screws of every two adjacent subsidiary fixtures form an included angel of 90 degrees; and the center lines of the screws of every two opposite subsidiary fixtures coincide with each other. By using the invention, the base can be adjusted in two degrees of freedom relative to the vibration table top, and the coincided centers of gravity of both a large test piece arranged on the base and the vibration table fixture can coincide with the axis of the vibration table when the large test piece is under the vibration test. The invention can ensure that the center of gravity of the test piece passes by the center of the vibration table top to reduce the influence of the overturning moment and can realize the rigid connection of the test piece and the vibration table top or the vibration table fixture.

The invention discloses application of a DNA target sequence of schistosoma japonicum retrotransposon in schistosomiasis diagnosis and particularly provides retrotransposon shown by any one nucleotide sequence selected from SEQ ID NO:1-25 and can be used as a detection marker for schistosomiasis. The invention also provides a pair of primers for specifically amplifying the retrotransposon sequence.

The invention belongs to the field of identification of Chinese medicinal materials, relating to a primer pair, kit and method and application thereof for identifying true and false of cordycepssinensis.In particular, the invention relates to the primer pair for identifying true and false of cordycepssinensis, which comprises CC-2F:5'-ATTAAGTCGTGGAAATG-3', CC-2R:5'-GATCAGGAATAGTGGGA-3', the kit thereof and the method for identifying the authenticity of the cordycepssinensis by PCR amplification by using the primer pair.The method has strong specificity and wider detection range.The invention also relates to the method for identifying the true and false of the cordycepssinensis by using the double primer pairs CC-2F/ CC-2R and DCF4/ DCR4 through PCR amplification, the amplification effect of the method is more obvious, the identification method is stable and reliable, and the identification range is wide.

The invention belongs to the technical field of molecular biology and virology and particularly relates to a universal test primer for viruses in a tobamovirus and a method for the universal test primer. Primer pair sequences of the universal test primer for the viruses in the tobamovirus are a forward primer TobamodF:5'-TKGAYGGNGTBCCNGGNTGYGG-3', and a reverse primer TobamodR:5'-ACNGAVTBNABCTGTAATTGCTAT-3. The test primer is designed on the basis of 32 conserved region sequences of a virus genome in the tobamovirus, amplification effect verification is carried out on the pair of primers by using seven viruses in the tobamovirus, and the amplification effects all are relatively good, thereby building universal test primer and method capable of testing at least seven viruses in the tobamovirus. The universal test primer has the characteristics of being high in testing efficiency, high in sensitivity and low in cost.

The invention discloses primers, probe and method for rapidly distinguishing an HP-PRRS (High pathogenic porcine reproductive and respiratory syndrome) vaccine strain GDr180 from an HP-PRRS wild strain. The method combines a real-time PCR (Polymerase Chain Reaction) technology with an MCA (Melting Curve Analysis) technology, and according to a difference of Tm values of a melting curve, the strain GDr180 and the wild strain are identified; the primers, the probe and the method are simple to operate, i.e. only the probe needs to be added before a PCR reaction; a detection speed is high and flux is high, i.e. the entire operating process only needs 3 hours, and time required for parting is greatly shortened; cost is relatively low, i.e. the identifying and detecting aims can be fulfilled only by one common probe; accuracy is high, specificity is good, repeatability is good, analysis can be accurately and rapidly carried out under the high flux, and the primers, the probe and the method are beneficial to popularization and application in clinical practice.