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42 results about "V region" patented technology

Method for carrying out high-throughput sequencing on TCR (T cell receptor) or BCR (B cell receptor) and method for correcting multiplex PCR (polymerase chain reaction) primer deviation by utilizing tag sequences

The invention provides a method for carrying out high-throughput sequencing on a TCR (T cell receptor) or a BCR (B cell receptor). The method is characterized by designing upstream primers according to gene features of a V region of the TCR or the BCR and designing downstream primers according to gene features of a C region or a J region of the TCR or the BCR and obtaining sequences of the of the TCR or the BCR in combination with the multiplex PCR (polymerase chain reaction) technology and high-throughput sequencing, thus analyzing the rearrangement information of the TCR or the BCR. Compared with 25-30 cycles of existing multiplex PCR, two cycles of the multiplex PCR technology provided by the invention can conduce to greatly reducing the sequencing errors caused by primer amplification preference. Besides, the invention also provides a method for correcting multiplex PCR (polymerase chain reaction) primer deviation by utilizing DNA (deoxyribonucleic acid) tag sequences, thus further reducing the sequencing errors caused by primer amplification preference and intrinsic sequencing errors of high-throughput sequencing.
Owner:SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA +1

Specific recognition sequence based T cell receptor high-throughput sequencing library construction and sequencing data analysis method

PendingCN111363783AComprehensive and Accurate Diversity InformationLower preferenceMicrobiological testing/measurementSequence analysisA-DNAV region
The present invention discloses a specific recognition sequence based T cell receptor high-throughput sequencing library construction and sequencing data analysis method. According to the method, a specific reverse transcription primer is designed for an mRNA sequence of a C region of a TCR constant region, cDNA is obtained by reverse transcription, and the 3' end of the cDNA is connected with a library construction linker with a specific recognition sequence; then the linker with the specific recognition sequence is added by using a splint connection method, and a TCR gene rearrangement sequence is amplified under the action of DNA polymerase by utilizing a gene specific primer with a tag; and finally a DNA library is added with a sequencing linker by PCR amplification to prepare a high-throughput sequencing library, and the high-throughput sequencing library is used for sequencing. The TCR gene diversity is comprehensively analyzed by bioinformatics, and a rearrangement rule of TCR genes including J region, D region and V region genes can be accurately obtained. The method is high in library construction efficiency, few in library construction step, low in required RNA initial quantity and low in library construction cost.
Owner:武汉康测科技有限公司

Multiple primer group and method for constructing human B cell immune repertoire with same on basis of high-throughput sequencing

ActiveCN109593758AReduce the cumbersomeness of the construction stepsReduce cumbersomeMicrobiological testing/measurementLibrary creationHuman b cellRNA
The invention discloses a multiple primer group and a method for constructing a human B cell immune repertoire with the same on the basis of high-throughput sequencing and belongs to the field of molecular biological detection. The multiple primer group consists of a group of upstream primers and a group of downstream primers, and the upstream primers are formed by connecting a joint 1, a primer bar code 1, a sequencing primer 1 and specific primers V1-V18 designed for V regions (variable regions) in series; the downstream primers are formed by connecting a joint 2, a primer bar code 2, a sequencing primer 2 and specific primers Va, Vd, Ve, Vg and Vm designed for conserved sequences of gene C regions (constant regions) of IgA, IgD, IgE, IgG and IgM in series; sequences of the primer bar code 1 and the primer bar code 2 are different while the sequences of the sequencing primer 1 and the sequencing primer 2 are the same. By means of the method, a BCR (B cell receptor) immune repertoirecan be constructed on the basis of a DNA sample or an RNA sample, and diversity information of BCRs can be completely covered. The construction efficiency is high and the cost is low.
Owner:SHANDONG ACV BIOTECH CO LTD

Variable region sequence library construction kit and sequencing method of variable region sequence

The invention belongs to the technical field of biology, and particularly relates to a variable region sequence library construction kit and a sequencing method of a variable region sequence. The variable region sequence library construction kit comprises a first primer group, primers of first connectors and a second primer group, the first primer group comprises first primers which specifically recognize all subtype coding sequences in a J region, each first primer comprises a nucleotide sequence, a proofreading random section and the first connector which are connected successively, the nucleotide sequences specifically recognize the coding sequences in the J region, the second primer group comprises successively connected second primers which specifically recognize all subtype coding sequences in a V region, and each second primer comprises a nucleotide sequence which specifically recognizes the coding sequences in the V region and a second connector. The variable region sequence library construction kit and the sequencing method of the variable region sequence are used for overcoming the technical defect that an existing TCR (T cell receptor) or BCR (B cell receptor) variable region sequence library construction technology is low in amplification efficiency, biased in amplification and prone to losing sequence information of a variable region sequence library.
Owner:GUANGZHOU HOMEY HEALTH TECH
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