An SNP marker for the detection of Rhd-negative phenotypes
A labeling, negative technology, applied in recombinant DNA technology, DNA/RNA fragments, microbial determination/inspection, etc., can solve problems such as inability to judge, easy to miss, and quality interference of samples
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Embodiment 1
[0023] Example 1: Screening of SNP markers
[0024] 1. Extraction of genomic DNA from peripheral blood:
[0025] In compliance with the relevant national policies and regulations, and on the basis of the consent of the sampling subjects, draw 2-5mL of peripheral venous blood from RhD-negative or mutant blood donors, put them into EDTA anticoagulant tubes, and store them at -80°C for later use; frozen EDTA anticoagulant blood After the coagulated blood melts at room temperature, take 500 μL and put it in a centrifuge tube, add an equal volume of TE (pH 8.0), mix well, centrifuge at 10,000 rpm at 4°C for 10 minutes, and discard the supernatant.
[0026] Add 180 μL TE, 20 μL SDS (10%), and 8 μL proteinase K (10 mg / ml) to mix well, and place in a 37° C. water bath overnight. Remove the sample from the water bath and briefly centrifuge to pellet the sample. Add an equal volume of Tris-saturated phenol (about 300 μL) to the reaction tube, mix thoroughly, centrifuge at 10,000 rpm f...
Embodiment 2
[0040] Example 2: Genetic Validation of Parents of a Proband with a SNP Mutation
[0041] 1. Extraction of genomic DNA from peripheral blood:
[0042] In compliance with the relevant national policies and regulations, and on the basis of the consent of the sampling subjects, draw 2-5mL of peripheral venous blood from RhD-negative or mutant blood donors, put them into EDTA anticoagulant tubes, and store them at -80°C for later use; frozen EDTA anticoagulant blood After the coagulated blood melts at room temperature, take 500 μL and put it in a centrifuge tube, add an equal volume of TE (pH 8.0), mix well, centrifuge at 10,000 rpm at 4°C for 10 minutes, and discard the supernatant.
[0043] Add 180 μL TE, 20 μL SDS (10%), and 8 μL proteinase K (10 mg / ml) to mix well, and place in a 37° C. water bath overnight. Remove the sample from the water bath and briefly centrifuge to pellet the sample. Add an equal volume of Tris-saturated phenol (about 300 μL) to the reaction tube, mix ...
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