Application of snp site and its primers, probes and detection kits
A site and reagent technology, which can be used in the determination/inspection of microorganisms, recombinant DNA technology, biochemical equipment and methods, etc., and can solve problems such as low accuracy and sensitivity, and impact on the accuracy of results
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Embodiment 1
[0126] Example 1 Using the kit provided by the present invention to assess the risk of graft injury and rejection after renal transplantation Extraction of total free DNA in plasma
[0127] 1. Using Streck non-invasive blood collection tubes to collect 10ml of blood from a recipient 2 weeks after kidney transplantation;
[0128] 2. Transfer the blood collected by the Streck non-invasive blood collection tube to a 15ml centrifuge tube, and centrifuge at 2000g at 4°C for 10min at low temperature; take about 4-6ml of the upper plasma in a new 15ml centrifuge tube, and continue to centrifuge at 4000g at 4°C for 10min at low temperature; take the upper plasma 4-6ml in a new 15ml centrifuge tube;
[0129] 3. Take 4ml of plasma, use QIAamp Circulating Nucleic Acid Kit from QIAGEN to extract the total free DNA, and adjust the DNA concentration to 10-40ng / ul.
[0130] Preparation of Digital PCR Reaction Droplets
[0131] 1. Configure a digital PCR reaction system, prepare digital PCR...
Embodiment 2
[0162] 2 ml of blood from the recipient 2 weeks after the kidney transplantation in Example 1 was collected using an EDTA blood collection tube, and routine renal function testing was performed. The results are shown in Table 5.
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