Primer composition for amplifying coding sequence of immunoglobulin heavy chain CDR3 and use thereof

An immunoglobulin and primer composition technology, which is applied in the field of primer compositions for amplifying the coding sequence of immunoglobulin heavy chain CDR3, and can solve problems such as the need for improvement of CDR3

Active Publication Date: 2013-07-03
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 composition for amplifying coding sequence of immunoglobulin heavy chain CDR3 and use thereof
  • Primer composition for amplifying coding sequence of immunoglobulin heavy chain CDR3 and use thereof
  • Primer composition for amplifying coding sequence of immunoglobulin heavy chain CDR3 and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] 1. Isolation of Human Peripheral Blood Mononuclear Cells

[0068] 1) Take 10mL of fresh peripheral blood from a healthy person, pour it into a sterile test tube containing anticoagulant and shake it well, and add an equal amount of sterile PBS to shake it well.

[0069] 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.

[0070] 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.

[0071] 4) Add 4 times the volu...

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PUM

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Abstract

The invention relates to a primer composition for amplifying the coding sequence of immunoglobulin heavy chain CDR3, a method for enriching the coding sequence of immunoglobulin heavy chain CDR3, a method for constructing the sequencing library of the coding sequence of immunoglobulin heavy chain CDR3, a method for determining the sequence information of the coding sequence of immunoglobulin heavy chain CDR3, a method for determining individual immune status, and a system for determining individual immune status. The primer composition comprises a first primer group and a second primer group, wherein the first primer group is composed of at least one V-region primer, and each of the at least one V-region primer comprises a sequence complementary with at least one V-gene segment; and the second primer group is composed of at least one J-region primer, and each of the at least one J-region primer comprises a sequence complementary with at least one J-gene segment. By virtue of the primer composition, the coding sequence of immunoglobulin heavy chain CDR3 can be effectively enriched, thus providing a convenient tool for deep researches on CDR3.

Description

technical field [0001] The present invention relates to the field of biotechnology. Specifically, the present invention relates to a primer composition for amplifying an immunoglobulin heavy chain CDR3 coding sequence and uses thereof. More specifically, the present invention relates to a primer composition for amplifying an immunoglobulin heavy chain CDR3 coding sequence, a method for enriching an immunoglobulin heavy chain CDR3 coding sequence, and a method for constructing a sequencing library of an immunoglobulin heavy chain CDR3 coding sequence The method, the method for determining the sequence information of the immunoglobulin heavy chain CDR3 coding sequence, the method for determining the immune status of an individual, and the 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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IPC IPC(8): C12N15/11C12N15/13C12N15/10C12Q1/68C40B40/08
Inventor 洪雪玉刘晓曾晓静苏政张俊杰王俊
Owner BGI SHENZHEN CO LTD
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