Primer combination for amplifying alpha chain CDR3 (complementary determining region 3) coding sequences of T cell receptor and use of primer combination

A technology of primer composition and cell receptor, which is applied in the biological field and can solve problems that need to be improved

Active Publication Date: 2013-09-11
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 the coding seque...

Method used

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  • Primer combination for amplifying alpha chain CDR3 (complementary determining region 3) coding sequences of T cell receptor and use of primer combination
  • Primer combination for amplifying alpha chain CDR3 (complementary determining region 3) coding sequences of T cell receptor and use of primer combination
  • Primer combination for amplifying alpha chain CDR3 (complementary determining region 3) coding sequences of T cell receptor and use of primer combination

Examples

Experimental program
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Embodiment 1

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

[0068] 1) Take 5ml of fresh peripheral blood from a healthy person in a 15ml centrifuge tube, add 5ml of PBS, mix well, then slowly add it to a 50ml centrifuge tube containing 6ml of mononuclear cell separation solution against the tube wall, 1,800rpm Centrifuge for 15 minutes.

[0069] 2) Aspirate the mononuclear cell layer into a new 15ml centrifuge tube, add three times the volume of PBS, mix gently, centrifuge at 1,800rpm for 10min, discard the supernatant to obtain a precipitate.

[0070] 3) The above pellet was resuspended with 1ml of PBS, then transferred to a new 1.5ml EP tube, centrifuged at 1,800rpm for 10min, and the supernatant was discarded to obtain the pellet.

[0071] 4) The pellet obtained in the above steps was resuspended with 100 μl of PBS, and the monocyte extraction was completed.

[0072] 2. Nucleic acid sample extraction

[0073] Utilize DNA extraction kit (Qiagen) to extract human per...

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Abstract

The invention relates to a primer combination for amplifying alpha chain CDR3 (complementary determining region 3) coding sequences of a T cell receptor and a use of the primer combination. The primer combination comprises a first primer set comprising at least one V region primer each of which comprises a sequence complementary with at least one V gene segment, and a second primer set comprising at least one J region primer each of which comprises a sequence complementary with at least one J gene segment. By virtue of the primer combination, the alpha chain CDR3 coding sequences of the T cell receptor can be effectively enriched, so that a convenient tool is provided for in-depth study of the alpha chain CDR3 of the T cell receptor.

Description

technical field [0001] The present invention relates to the field of biotechnology. Specifically, the present invention relates to a primer composition for amplifying T cell receptor alpha chain CDR3 coding sequence and its use. More specifically, the present invention relates to a primer composition for amplifying the coding sequence of T cell receptor α chain CDR3, a method for enriching the coding sequence of T cell receptor α chain CDR3, and a method for constructing the coding sequence of T cell receptor α chain CDR3 A method for sequencing a library, a method for determining the sequence information of the T cell receptor alpha chain CDR3 coding sequence, a method for determining the immune status of an individual and a system for determining the immune status of an individual. Background technique [0002] The immune system is an important system for the body to resist the invasion of pathogenic bacteria and regulate immunity, so it is of great significance to study ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/12C12N15/10C40B40/08C40B50/06C12Q1/68C12M1/34
Inventor 张瑞芳刘晓洪雪玉郑春婷苏政张俊杰
Owner BGI SHENZHEN CO LTD
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