Treatment method of fluorescent PCR amplification sample and kit
A treatment method and a sample treatment solution technology, applied in the field of biomedicine, can solve the problems of cumbersome operation, easy cross-contamination, large DNA loss, etc., and achieve the effect of reducing loss and wide application range
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Embodiment 1
[0022] Embodiment 1: Blood samples are processed by the method of the present invention
[0023] (1) Experimental material: EDTA anticoagulant blood
[0024] (2) Experimental method:
[0025] A. Gently mix the EDTA anticoagulant blood, take 1-10μL and place it at the bottom of a 1.5mL centrifuge tube;
[0026] B. Add 200 μL of sample processing solution;
[0027] C. After vortexing and mixing, briefly centrifuge;
[0028] D. Put it in the centrifuge tube rack and set it aside.
Embodiment 2
[0029] Example 2: Evaluation of the processing effect of the blood sample processed by the method of the present invention
[0030] (1) Experimental materials: 20 parts of EDTA anticoagulant blood
[0031] (2) Experimental method:
[0032] A. Take 200 μL of 20 EDTA anticoagulated blood samples, use domestic blood genomic DNA extraction kit to extract genomic DNA, and measure DNA concentration by spectrophotometer;
[0033] B. The same 20 EDTA anticoagulant blood sample, take 5 μ L respectively, and mix with 200 μ L of the sample treatment solution provided by the present invention respectively;
[0034] C. get each 2 μ L of the genomic DNA extracted by the kit and the processed sample of the present invention, and add to the PCR tube respectively;
[0035] D. add IL28b fluorescent PCR reaction liquid, wherein reaction liquid comprises the following components of following concentration or content: 10mM Tris-HCl (pH8.3), 50mM KCl, 1.5mM MgCl 2 , 200μM dNTPs, 200nM upstream a...
Embodiment 3
[0043] Example 3: Detection of IL28b gene rs12979860 polymorphism
[0044] (1) Experimental materials: 20 parts of EDTA anticoagulant blood
[0045] (2) Experimental method:
[0046] A. 20 parts of EDTA anticoagulated blood samples, each take 5 μ L, mix with 200 μ L of the sample treatment solution provided by the present invention respectively;
[0047] B. Take 2 μL of each processed sample and add it to a PCR tube;
[0048] C. add IL28b fluorescence PCR reaction liquid, wherein reaction liquid comprises the following components of following concentration or content: 10mM Tris-HCl (pH8.3), 50mM KCl, 1.5mM MgCl 2 , 200μM dNTPs, 200nM upstream and downstream primers, 200nM dual probe (type C and type T), 0.5U Taq DNA polymerase.
[0049] D. Carry out amplification detection in the fluorescent PCR instrument that model is ABI 7500, described fluorescent PCR amplification program is as follows:
[0050] Step 1: 60°C, 30s; cycle 1 time, collect fluorescence;
[0051] Step 2: 95...
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