Kit, device and method for increasing fetal free dna concentration in maternal peripheral blood
A peripheral blood, high-concentration technology, used in biochemical cleaning devices, biochemical equipment and methods, enzymology/microbiology devices, etc., can solve the problems of missed detection of positive samples, false negatives, etc. Concentration, cost reduction effect
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Embodiment 1
[0065] Embodiment 1: A kind of kit that improves fetal cell-free DNA concentration in maternal peripheral blood
[0066] The kit consists of free DNA extraction reagents, library building reagents and free DNA enrichment reagents, among which the free DNA enrichment reagents can enrich free DNA fragments at any point or any segment of free DNA with a fragment length not exceeding 160bp .
[0067] Specifically, the free DNA extraction reagent is a magnetic bead extraction reagent, including proteinase K, lysis buffer, cleaning solution 1, cleaning solution 2, and eluent.
[0068] Library building reagents are library building reagents for fetal chromosomal aneuploidy, including: end repair buffer, end repair enzyme, DNA ligase, DNA ligase, PCR amplification reagent, PCR primers, TE solution, P1 adapter, label Sequences 1-32; Ion Plus Fragment Library Ki (Life Technology) or New England Biolabs, and related library construction reagents from Enzymatics can also be used. In thi...
Embodiment 2
[0117] Embodiment 2: A kind of test kit that improves fetal cell-free DNA concentration in maternal peripheral blood
[0118] The composition of the kit is the same as in Example 1.
[0119] Samples and their processing: The samples come from 10ml maternal venous blood samples of Beijing Boao Medical Laboratory, numbered "RD13120110", "RD13120111", "RD13120112" and "RD13120113". These samples were all collected from the peripheral blood of pregnant women who were pregnant with male fetuses. Among them, RD13120112 was a T21-positive sample, and the other three samples were confirmed as normal fetuses by clinical follow-up. Blood samples were centrifuged in two steps to obtain blood cell-depleted plasma, which could be used for subsequent experiments or frozen at -80°C for future use.
[0120]Extract plasma free DNA: select the serum / plasma free DNA magnetic bead extraction kit (product number S10020) produced by Boao Muhua Company for DNA extraction, extract three copies of DN...
Embodiment 3
[0147] Embodiment 3: A kind of device that improves fetal cell-free DNA concentration in maternal peripheral blood
[0148] Obtain short fragments of free DNA <160bp in a certain section or a certain point in maternal peripheral blood by physical methods, and the devices for separating short fragments of free DNA by physical methods are gel electrophoresis, prefabricated gel electrophoresis, etc., including electrophoresis tanks, gels, DNAladder, electrophoresis platform and operating system.
[0149] The device includes a peripheral blood free DNA separation device, a free DNA library construction device, a library purification device and a free DNA enrichment device for enriching free DNA fragments at any point or segment of free DNA whose fragment length does not exceed 160 bp. in:
[0150] The peripheral blood cell-free DNA separation device is a magnetic bead extraction kit, including: proteinase K, lysis buffer, cleaning solution 1, cleaning solution 2, and eluent;
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