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Primer set used for amplifying immunoglobulin light-chain CDR3 sequences, and its uses

An immunoglobulin and primer set technology, which can be used in DNA/RNA fragments, DNA preparation, recombinant DNA technology, etc., and can solve problems such as CDR3 needs to be improved.

Active Publication Date: 2015-06-17
BGI SHENZHEN CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the current research on CDR3 still needs to be improved

Method used

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  • Primer set used for amplifying immunoglobulin light-chain CDR3 sequences, and its uses
  • Primer set used for amplifying immunoglobulin light-chain CDR3 sequences, and its uses
  • Primer set used for amplifying immunoglobulin light-chain CDR3 sequences, and its uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] 1. Isolation of Human Peripheral Blood Mononuclear Cells

[0069] 1) Take 10 mL of fresh peripheral blood from a healthy person, pour it into a sterile test tube containing anticoagulant and shake it well, then add the same amount of sterile PBS and shake it well, the sample name is ZXJ.

[0070] 2) Take an appropriate volume of Percoll layering solution (the volume ratio of peripheral blood and layering solution is 2:1 to 3:1) in a test tube, and slowly add the diluted blood to the layering solution surface to form a clear interface , placed in a horizontal centrifuge, 2000r / m, 20 minutes.

[0071] 3) After centrifugation, it can be seen that there are four layers from the bottom of the centrifuge tube to the liquid surface, which are red blood cell and granulocyte layer, stratified liquid layer, mononuclear cell layer, and plasma layer. Use a pipette to insert under the liquid surface, draw the mononuclear cell layer, and place it in another sterile test tube.

[00...

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PUM

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Abstract

The invention relates to a primer set used for amplifying immunoglobulin light-chain CDR3 sequences, a kit and methods thereof, a method for enriching the immunoglobulin light-chain CDR3 sequences, a method for constructing a sequencing library of the immunoglobulin light-chain CDR3 sequences, a method for determining the sequence information of the immunoglobulin light-chain CDR3 sequences, a method for determining individual immunization states, and a system for determining the individual immunization states. The primer set comprises: a forward primer group, wherein the forward primer group is composed of at least one V region primer, and each of the at least V region primer includes a sequence complementary with at least one V gene fragment; and a backward primer group, wherein the backward primer group is composed of at least one J region primer, and each of the at least J region primer includes a sequence complementary with at least one J gene fragment. The primer set can effectively enrich the immunoglobulin light-chain CDR3 sequences, so the primer set provides a convenient tool for deep-going researches on CDR3.

Description

technical field [0001] The present invention relates to the field of biotechnology. In particular, the present invention relates to primer sets and uses thereof for amplifying immunoglobulin light chain CDR3 sequences. More specifically, the present invention relates to a primer set for amplifying immunoglobulin light chain CDR3 sequences, a method for amplifying immunoglobulin light chain CDR3, a method for enriching immunoglobulin light chain CDR3 sequences, and constructing immunoglobulin light chain CDR3 sequences. A method for sequencing libraries of chain CDR3 sequences, a method for determining the sequence information of immunoglobulin light chain CDR3 sequences, a method for determining the immune status of an individual, and a system for determining the immune status of an individual. Background technique [0002] Immunoglobulins, T-cell receptors and HLA (Human Leukocyte Antigens) are the most active molecules in the human genome, determining and reflecting the i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/68C40B50/06
Inventor 不公告发明人
Owner BGI SHENZHEN CO LTD
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