A primer set, method and kit for detecting fetal triploid
A technology of triploid and primer set, which is applied in biochemical equipment and methods, recombinant DNA technology, and microbial measurement/inspection, etc. It can solve the problems of high requirements for PCR reaction conditions, large number of primers, and low efficiency, so as to improve Population quality, process and data analysis are simple and cost-effective
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[0050] (1) Separation and extraction of cfDNA:
[0051] Collect 10ml of venous blood from pregnant women (12-24 weeks of pregnancy) into vacuum blood collection tubes (EDTA anticoagulant), centrifuge at 1600g for 10 minutes, and centrifuge at 16000g for 10 minutes to obtain about 4ml of plasma, and use a free DNA extraction kit to separate free DNA in the plasma , to get 15ng cfDNA.
[0052] (2) Primer design:
[0053] For human chromosomes 13, 18, and 21, 300 pairs of sequence-specific primers were designed respectively. The sequence-specific primer has the following characteristics: (1) Each site of the primer is composed of two consecutively arranged specific position oligos dependent on peripheral blood free DNA (cfDNA) through an intermediate oligonucleotide bridge Forming a PCR template; (2) the primers have a unique sequence on the target chromosome; (3) the position-specific oligonucleotides have the same annealing temperature; (4) the primers are common to the seque...
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