59results about How to "Inhibition of nonspecific amplification" patented technology

The invention relates to a preparation method?of a hot start DNA polymerase,?comprising the following steps:?obtaining a?single stranded DNA binding protein, a Taq DNA polymerasemonoclonal antibody?and?a Taq DNA polymerase; and mixing the?single stranded DNA binding protein, the Taq DNA polymerase?monoclonal antibody?and the Taq DNA polymerase in accordance with?aproportion. The contents of the?single stranded DNA binding protein, the Taq DNA polymerasemonoclonal antibody?and the Taq DNA polymerase are all 20-40%?respectively. The invention also relates to the hot start DNA polymerase prepared by the above method and an application thereof. The hot start DNA polymerase provided by the invention can also be used to solve problems which appear easily in hot start PCR technology such as DNA damage, product contamination?and bad effect of hot start caused by incomplete sealing.

The invention relates to a B-raf gene mutation detection kit. The B-raf gene mutation detection kit comprises quality-control primer probe internal-standard mixed liquor and detection primer probe internal-standard mixed liquor, wherein the quality-control primer probe internal-standard mixed liquor comprises a quality-control primer pair, a B-raf gene specific probe, an internal-standard primer pair, an internal-standard specific probe and an internal-standard template, and the detection primer probe internal-standard mixed liquor comprises a B-raf gene V600E mutation detection specific primer pair, a B-raf gene specific probe, an amplification blocking nucleic acid sequence, an internal-standard primer pair, an internal-standard specific probe and an internal-standard template. The B-raf gene mutation detection kit has the advantages that an amplification refractory mutation system is combined with a wild type amplification blocking nucleic acid sequence with a phosphorylated terminal, deoxyinosine is introduced into detection of a B-raf gene V600E mutation detection specific ARMS (amplification refractory mutation system) forward primer to enable quality-control PCR (polymerase chain reaction) and detection PCR to be performed for detection of samples parallelly, and each reaction system can have an internal-standard system capable of effectively avoiding false negative and intra-assay variation; the B-raf gene mutation detection kit is low in cost, high in sensitivity and more capable of controlling intra-assay and inter-assay variation.

The invention discloses probes, primers and kits for detecting seven kinds of mutations in human NRAS genes, wherein there are seven sets of primers and probes from NM1 to NM7, and the mutation primers in each group can combine with the conserved sequence of the NRAS gene, and the mutation ARMS primers It can specifically bind to the corresponding mutant sequence and amplify the mutant sequence; the detection probe can bind to the amplified fragment, and the blocking probe can specifically bind to the wild-type sequence corresponding to the mutation site, inhibiting the non-specific amplification of the wild-type . The present invention adopts specific mutation primer and probe blocking technology, can accurately detect 7 kinds of mutation types of NRAS gene; the kit of the real-time fluorescent PCR amplification reaction system established is convenient for rapid detection of NRAS gene mutation, and the operation is simple and the result is easy Read; the detection method has high sensitivity, and 50 copies of mutations can be detected stably; the specificity is good, 20ng of wild-type genomic DNA has no non-specific amplification, and the detection ability is as high as 1%.

The invention discloses a triple fluorescence PCR primer set for detecting pepper transgenic ingredients. The triple fluorescence PCR primer set comprises a triple primer set A for CaATL1, CaMV35S and NPT II genes and a triple primer set B for NOS, CMV and TMV genes and is specifically shown as Seq. ID No.1 to Seq. ID No.12. The invention belongs to the technical field of genetic engineering detection; all the primers and probes cannot mutually interfere; a specific target sequence is amplified and excites a fluorescence signal, the non-target sequence is not amplified and generates no fluorescence signal and accurate detection for the pepper transgenic ingredients can be realized; the triple fluorescence PCR primer set has the advantages of high specificity, low standard error, short detection time, capability of saving reagent cost and the like, and is suitable for the detection for the transgenic ingredients of the pepper product.