High-sensitivity detection method applied to forensic mixed samples
A technology of sensitive detection and detection method, which is applied in the field of forensic application research and can solve the problems of sensitive and accurate typing of samples that cannot be mixed.
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Embodiment 1
[0086]Example 1 A high sensitive detection technology applied to a forensic mixed projection material
[0087]1. Sample preparation: The inspection material is from the complex cytokines from the public security criminal case, and the extraction of DNA is used in the extraction of magnetic bead method. Specific method according to the relevant standards of the public security system (GA383-2014), this example Pick 3 samples for testing, sample name sample 1, sample 2, sample 3;
[0088]2. Detection method: The detection method is detected in accordance with the detection method of the present invention;
[0089]3. Data analysis: Electrophoresis in genetic analyzers, injection time is 10 seconds, the injection voltage is 3kV; software analysis, set the peak high threshold is 50Ruf;
[0090]4. Results analysis: The conventional control method has not been complete sample 1, sample 2, sample 3 fragmentation, specific detection sample repeated fragment repeated number and amplification fragment siz...
Embodiment 2
[0100]Example 2 A high sensitive detection technology applied to a forensic mixed projection material
[0101]1. Sample: Example 1 Sample 1 gradient dilution liquid sensitivity detection;
[0102]2. Method: The nucleic acid extracted with Qubit 3.0, dilute the sample 1 gradient to 1 ng / μL, 0.5 ng / μL, 0.25 ng / μL, 0.1 ng / μL, 0.05 ng / μL, 0.025 ng / μL, 0.01 ng / μL is detected in accordance with the detection method of the present invention;
[0103]3. Data analysis: Electrophoresis in genetic analyzers, injection time is 10 seconds, the injection voltage is 3kV; software analysis, set the peak high threshold is 50Ruf;
[0104]4. Results: Conventional Control Detection Method Example 1 Sample 1 Gradient Diluent: 1 ng and 0,5 ng All of the 8 alleles were detected; 4 alleles were detected at 0.25 ng, and the detection rate was 50%; 0.1 ng 2 alleles were detected, the detection rate was 25%; the adjustment threshold to 20RUF, 0.01 ng detected 2 alleles, and the fluorescent signal was 41Ruf,...
Embodiment 3
[0107]Example 3 A high sensitive detection technology applied to a forensic mixed projection
[0108]Detection of mixed sample low abundance target sequence;
[0109]1. Sample Preparation: Use the sample 1 and sample 2 of Example 1 to mix the DNA of the sample 1 and the sample 2, and the target target proportional to the sample in the sample is: 1%, 0.1%, 0.05%, respectively. 0.01%, 0.005%, 0.001%, and diluted with water to a final concentration of 10 ng / μL, and the total volume of mixed sample was 60 μl. Take 1 μL of the mixed in addition to the test;
[0110]2. Determination of the mixture: Sample 1: Low abundance target is 1%; mixed 2: low abundance target is 0.5%; mixed 3: low abundance target is 0.1%; mixed 4: low abundance The degree target is 0.05%; mixed 5: low abundance target is 0.01%; mixed 6: low abundance target is 0.005%; mixed 7: low abundance target is 0.001%; the detection method of the present invention ;
[0111]3. Data analysis: Electrophoresis in genetic analyzers, the ...
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