Eight-color system STR (short tandem repeat) site multiplex amplification kit aiming at difficult detection material and application of eight-color system STR site multiplex amplification kit
A compound amplification and kit technology, applied in the field of molecular biology, can solve the problems such as the inability to meet the requirements of higher identification ability, the limited number of fluorescence channels of genetic analyzers, poor complementarity and versatility, etc., and achieve amplification of fragments. The effect of short, good polymorphism and short amplification time
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Embodiment 1
[0033] 30pg and 0.5ng standard product 9948 were detected using the kit of the present invention.
[0034] The kit of the present invention consists of the following: primer mixture, 1mL; PCR buffer Unique Mix, 2mL; sdH 2 O, 1.95 mL; PCR enhancer.
[0035] The composition of the PCR buffer Unique Mix is: hot start Taq enzyme 0.2U / μL~0.5U / μL, Tris-HCl 50mM~125mM with pH 8.25~8.4, dNTPs 5.0mM~7.5mM, KCl 50mM~125mM, BSA 2mg / ml~5mg / ml.
[0036] The optional types of PCR enhancers are: polyethylene glycol, Triton X-100, polyvinylpyrrolidone (PVP) or dithiothreitol, and one or two of them are selected and directly added to the PCR buffer Unique Mix.
[0037] 1. The operation steps are as follows:
[0038] 1.1 Amplification system
[0039] Component volume Unique Mix 10.0 μL 30pg / 0.5ng9948 1μL Primer set 5.0 μL wxya 2 o
Make up to 25 μL
[0040] 1.2 The amplification procedure is:
[0041]
[0042] 1.3. Fluorescent detection of...
Embodiment 2
[0047] A total of 24 samples of samples such as saliva spots, mouse swab samples, and old degraded samples in actual cases were detected using the kit of the present invention.
[0048] The kit of the present invention consists of the following: primer mixture, 1mL; PCR buffer Unique Mix, 2mL; sdH 2 O, 1.95 mL; PCR enhancer.
[0049] The composition of the PCR buffer Unique Mix is: hot start Taq enzyme 0.2U / μL~0.5U / μL, Tris-HCl 50mM~125mM with pH 8.25~8.4, dNTPs 5.0mM~7.5mM, KCl 50mM~125mM, BSA 2mg / ml~5mg / ml.
[0050] The optional types of PCR enhancers are: polyethylene glycol, Triton X-100, polyvinylpyrrolidone (PVP) or dithiothreitol, and one or two of them are selected and directly added to the PCR buffer Unique Mix.
[0051] 1. Sample preparation: extract by magnetic bead method, refer to "GA / T 383-2002 Forensic Science DNA Laboratory Inspection Specification" for specific methods.
[0052] 2. The operation steps are as follows:
[0053] 2.1 Amplification system
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