DNA library concentration measuring method for fetus chromosome aneuploid detection
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A technology for aneuploidy and concentration determination, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of poor practical operation effect, difficult to copy operation, unclear standards, etc., and improve labor efficiency , time saving, simple and convenient operation
Inactive Publication Date: 2018-10-12
CHANGSHA KINGMED MEDICAL DIAGNOSTICS INST
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[0008] The main problem solved by this application is to provide a DNA library concentration determination method in the detection of fetal chromosomal aneuploidy, which has simple operation, scientific process, clear standards, and good practical operation effect, so as to solve a detection method for fetal chromosomal aneuploidy The DNA library concentration determination method in China has high cost, unscientific process, unclear standards, poor practical operation effect, high operation difficulty, and difficult to replicate technical problems
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Embodiment 1
[0028] A DNA library concentration determination method in the detection of fetal chromosomal aneuploidy, comprising:
[0029] Step 1: Dilute the sample to be tested, prepare the standard, and store both at low temperature;
[0030] Step 2: Take the centrifuge tube, mark it on the cap, prepare the reaction mixture according to the reaction system in the table below, and prepare the system according to the amount required for each sample to do 2 repeated reactions. The reaction mixture includes quantitative After the amplification reagent, nuclease-free water, P1 primer and P2 primer are prepared, shake the reaction mixture for 10 seconds and centrifuge briefly to the bottom of the tube;
[0031] Step 3: Add the reaction mixture prepared in step 2 to each reaction tube hole of the reaction plate, add 2L of standard product to the standard product reaction wells, and set no template control reaction wells on each reaction plate at the same time , add 2uL nuclease-free water to ...
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Abstract
The invention discloses a DNA library concentration measuring method for fetus chromosome aneuploid detection. The method comprises the steps of 1, diluting a sample to be tested, preparing a standardproduct, and performing cryopreservation on the sample and the product; 2, taking a centrifugal tube, marking a tube cover, preparing a reaction mixture according to a reaction system in the table, wherein during preparation, according to a needed quantity preparing system for two-time repeated reactions of each sample, the reaction mixture comprises a quantified amplification regent, nuclear free water, a P1 primer and a P2 primer, after preparation is completed, the reaction mixture is oscillated, and instant centrifuging is performed till the tube bottom; 3, adding the reaction mixture prepared in step 2 into reaction tube holes of a reaction plate, and adding 2 L standard products into the standard product reaction holes; 4, sealing the reaction plate, centrifuging the reaction plate,loading the reaction plate on a fluorescent quantitation PCR cycler, the qPCR reaction begins to be executed after setting is performed, and a library concentration measuring result is obtained; operation is easy, the flow path is scientific, the standard is clear, and the practical operation effect is good.
Description
technical field [0001] The invention belongs to the field of non-invasive prenatal screening and detection, and specifically refers to a DNA library concentration determination method in the detection of fetal chromosome aneuploidy. Background technique [0002] Chromosomal aneuploidy refers to the increase or decrease in the number of one or several chromosomes in a cell compared to the normal 46 chromosomes in a human being, which is closely related to the morbidity and mortality in infants and young children. The incidence rate of chromosomal abnormalities in newborns is 1 / 160, of which trisomy 21 (Down syndrome), trisomy 18 (Edward syndrome) and trisomy 13 (Patau syndrome) are the three most common The incidence rates of chromosomal aneuploidy in newborns are 1 / 800-1 / 600, 1 / 7000-1 / 3500 and 1 / 6000-1 / 5000, respectively. Prenatal testing for chromosomal diseases mainly includes serological screening and amniocentesis. However, the accuracy of serological screening is low,...
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