Human chromosome 15q11-13 genetic variation detection kit and application thereof
A detection kit and a technology for genetic variation, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of lack of detection tools, etc., and achieve the effect of good typing map, balanced amplification, and avoiding pollution
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Embodiment 1
[0045] Example 1: Preparation of genomic DNA from human blood samples
[0046] Test samples were donated by volunteers with informed consent. According to medical routine, 1 mL of peripheral venous blood was collected and anticoagulated with EDTA. Genomic DNA was extracted using the Human Peripheral Blood Genome Extraction Kit from QIAGEN, with an elution volume of 100 microliters, quantified using a UV spectrometer, and the concentration of genomic DNA was diluted to 1 ng / μL.
Embodiment 2
[0047] Embodiment 2: PCR system preparation
[0048] Prepare the PCR reaction system according to the following system (the total reaction system is 20 μL):
[0049] 2×PCR reaction master mix 10μL
[0050] Primer mix 2 μL
[0051] Sample DNA template 1 μL
[0052] wxya 2 O 7μL
[0053] Take out the primer composition container and the PCR reaction mother solution container from the kit. According to the total number of reactions multiplied by the individual reaction requirements in the above reaction system, the required amount of PCR reaction mother solution, primer composition amount, PCR reaction auxiliary liquid amount and ddH 2 O amount, mix the above reagents evenly in a 1.5mL EP tube, divide and number each PCR reaction tube 19 μL, then add 1 μL of the sample DNA template according to the sample number, and mix again.
Embodiment 3
[0054] Embodiment 3: PCR reaction
[0055] ABI 9700 PCR instrument was used for PCR reaction.
[0056] The PCR conditions are as follows: 95°C for 10 minutes after starting, 95°C for 30 seconds, 60°C for 60 seconds, 72°C for 60 seconds, a total of 30 cycles, then 72°C for 60 minutes, and then 4°C for incubation.
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