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Primer group, kit and method special for Diego blood group gene sequencing

A gene sequencing and primer set technology, applied in biochemical equipment and methods, recombinant DNA technology, and microbial determination/examination, etc., can solve problems such as difficulty in determining Digeo blood group genes, and achieve the effect of assisting clinical diagnosis.

Pending Publication Date: 2021-11-12
江苏中济万泰生物医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved in the present invention is to overcome the difficulties in the determination of Digeo blood group genes due to the occurrence of gene mutations in the prior art, and provide a primer set, kit and method dedicated to the sequencing of Diego blood group genes

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0034] DNA extraction: Generally, 2ml of EDTA anticoagulated blood sample is taken, and the DNA is automatically extracted by the magnetic bead method, and the measured OD (260 / 280) = 1.9.

[0035] PCR amplification:

[0036] 1. Preparation of PCR working solution: Add 500 μl of water and 10 μl of Taq enzyme into the PCR buffer tube, mix and centrifuge.

[0037] 2. Preparation of PCR working solution and sample DNA mixture: add 1 microliter of forward and reverse sequencing primer (10pmoles) reaction tubes, 40mL of PCR buffer and 4mL of DNA for PCR reaction.

[0038] PCR amplification program:

[0039] step temperature and time cycle 1 96°C 6min 1cycle 2 96°C 15sec, 68°C 60sec 30cycles 3 96°C 20sec, 65°C 50sec, 72°C 45sec 10cycles 4 72℃ 2min 1cycle 5 Store at 4°C

[0040] Or PCR amplification procedure:

[0041] step temperature and time cycle 1 98℃ 7min 1cycle 2 98°C 10sec, 55°C 30sec, 72°C...

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PUM

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Abstract

The invention discloses a kit and a method special for Diego blood group gene sequencing. The kit and the method are characterized in that key segments of alleles containing mutation sites are optimally selected, and matched amplification reagents and purification procedures are added to realize Sanger method sequencing of target alleles. The primer group special for Diego blood group gene sequencing can solve the problems of antigen identification of Diego blood groups, judgment of difficult blood groups, discovery of new mutation sites, gene polymorphism detection and the like.

Description

technical field [0001] The invention relates to the technical field of gene diagnosis products, in particular to a primer set, a kit and a method specially used for sequencing the Diego blood type gene. Background technique [0002] The gene for the Diego blood group system is SLC4A1, which encodes a protein that is part of a family of anion exchangers expressed in the plasma membrane of red blood cells and is involved in the transport of carbon dioxide from tissues to the lungs as a chloride / bicarbonate exchanger. The SLC4A1 gene is located on chromosome 17q21.31 and contains 20 exons. [0003] There are currently 23 antigens found in the Diego blood group system, which have important clinical significance. Blood type incompatibility can cause hemolytic transfusion reactions and hemolytic disease of the newborn, and even endanger the lives of patients. Each antigen corresponds to a specific allele, which can be determined by detecting the mutation site. The existing tech...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6869C12N15/11
CPCC12Q1/6888C12Q1/6869C12Q2600/156C12Q2531/113C12Q2535/101C12Q2565/125
Inventor 高宏军陈玉平王亚恒陈芳芳
Owner 江苏中济万泰生物医药有限公司