124 results about "Hybridization capture" patented technology

The present invention provides methods of detecting chromosomal or genetic abnormalities associated with various diseases or with predisposition to various diseases. In particular, the present invention provides advanced methods of performing DNA hybridization, capture, and detection on solid support. Invention methods are useful for the detection, diagnosis, predicting response to therapy, detecting minimal residual disease, prognosis, or monitoring of disease treatment or progression of particular disease conditions such as cell proliferative disorders

The invention discloses a building method of a library for detecting non-small cell lung cancer gene mutation and a kit. The method includes: using tubular reaction to complete genome DNA breaking and connector connection, performing hybrid capture on connection products after amplification and non-small cell lung cancer related gene target area probes, and performing BGISEQ-500 / 1000 platform sequencing and data analysis to obtain mutation conditions. The method has the advantages that the experiment flow is optimized greatly by the tubular reaction, operation complexity and time are reduced, and the requirements on clinical sample initial amount are lowered; multiple genes and multiple sites can be detected in one step, point mutation, insertion and deletion, structural variation and copy number variation are covered, the detecting result is accurate and overcomes the defect that a PCR capture method cannot detect the structural variation in one step, and the effectiveness of the high-throughput sequencing applied to the detection of the non-small cell lung cancer gene mutation; the method is wide in coverage, high in cost performance, capable of providing a reference basis for the diagnosing, treatment and drug use performed by doctors, and the method is suitable for being popularized and used in a large-scale manner.

The invention provides a method for efficiently and rapidly separating a T-DNA insertion site flanking sequence. The method comprises: extracting the genomic DNA of an Agrobacterium-mediated transgenic plant; fragmenting the genomic DNA, carrying out DNA repairing, adding A, linking, carrying out fragment selection by using magnetic beads, carrying out library construction, carrying out hybridization capture on the constructed library and a T-DNA boundary sequence, and carrying out PCR enrichment; and carrying out high-throughput sequencing on the library, and carrying out bioinformatics analysis on the sequenced data so as to obtain the T-DNA insertion site. According to the present invention, the capture is performed with the target DNA, and the low-throughput sequencing is specially performed on the T-DNA boundary sequence; the disadvantages of low throughput, low efficiency and low success rate of the existing flanking sequence separation technology are overcome with the method of the present invention; and with the method of the present invention, the DNA capture and the second-generation sequencing technology are combined, the T-DNA boundary sequence is specially sequenced and the simple analysis is performed to obtain the T-DNA insertion site flanking sequence, and the method has characteristics of high efficiency, economy, and simple analysis.

The invention discloses a construction method of a fusion gene detection library based on an Ion Torrent sequencing platform and a fusion gene detection method. The construction method comprises the following steps that 1, fragmentation is conducted on sample DNA to obtain DNA fragments; 2, tail end repairing and basic group A adding are conducted on the DNA fragments; 3, joints with known sequences are connected; 4, PCR amplification is conducted to obtain a sample nucleic acid library; 5, the sample nucleic acid library captures DNA fragments of fusion genes and partner genes on the basis of probe hybridization of a target area capturing platform; 6, a product obtained in the step 5 is introduced into a sequencing joint sequence through PCR, and the fusion gene detection library is obtained. By means of the technical scheme, the problems that in the prior art, due to the fact that an RNA sample is not qualified, the Ion Torrent sequencing platform cannot detect the fusion genes and cannot detect the fusion types of unknown fusion breakpoints are solved.

The invention discloses a targeted trapping and sequencing kit for 285 lung cancer-related genes and application of the targeted trapping and sequencing kit. The kit is based on hybrid trapping of a probe, and can be used for preparing a high-throughput library of the lung cancer-related 285 genes and simultaneously detecting all variation loci on exons of the 285 genes, so that the disadvantagesof high cost, high time consumption, tedious operation and the like of conventional single-gene detection are overcome. Detection of a plurality of genes with a single small sample is implemented in the small sample for aspiration biopsy of lung cancer, and the clinical practice requirements of maximally using the small sample and obtaining a maximum amount of information are met. The kit is basedon an Ion Proton platform, has high accuracy and sensitivity, and has obvious advantages compared with a Sanger sequencing method, and a detection result in a residual DNA (deoxyribonucleic acid) sample of a paraffin sample is stable.

The invention provides a method for preparing a DNA library and specifically capturing and a kit, and the method is suitable for constructing the DNA library of cfDNA, white cell gDNA and DNA from a tissue sample as well as hybridizing and capturing a special target gene area. In the preparation method for the DNA library, terminal repair and 3' end A addition are performed in the same reaction system, and hybridization input is improved, and a blocking preparation in hybridizing and capturing is optimized, so that higher capturing efficiency, higher library variety and more uniform coverage degree are obtained, and therefore, the preparation method is especially suitable for constructing a trace sample library with low-abundance mutation, and is beneficial for realizing saving in technical process. The method provided by the invention can be used for conveniently realizing all variations such as mutation, deletion, insertion, fusion as well as detection on copy number amplification of a target gene, and overcomes the defects that target gene sequences are enriched through a PCR method on DNA level at present, mutation and deletion of the target gene only can be detected, and fusion cannot be detected.

The invention discloses a system capable of simultaneously detecting tumor targeting therapy related target spots and immunotherapy related TMB and MSI. The system comprises a library preparation reagent, a probe, a hybridization capture reagent and a data analysis system, wherein the library preparation reagent, the probe and the hybridization capture reagent can be used for sequencing a library,and the data analysis system can analyze results including data quality control information and statistical information, mutation site information and TMB values.

The invention belongs to the technical field of gene engineering, and particularly relates to a detection method of a low frequency mutated DNA fragment and a library building method. The technical problems that in the existing detection technique, the DNA fragment capturing efficiency is low, wrong mutation is introduced in the library building process, the detection limit is low, and the sensitivity is low are solved by adopting the hybrid capturing and molecular tagging technique and meanwhile combining with a special library building mode, the library specificity is improved, the false positive rate is lowered, and the template utilization rate is improved.

This invention is related to a method for rapidly detecting gene specific methylation through signal amplification using the epi-barcode after hybrid capture of the methylated DNA sequence.

Target-specific hybrid capture (TSHC) provides a nucleic acid detection method that is not only rapid and sensitive, but is also highly specific and capable of discriminating highly homologous nucleic acid target sequences. The method produces DNA-RNA hybrids which can be detected by a variety of methods.