Method for performing STR typing on forensic mixed DNA sample

A sample and biological sample technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of low suspects, limited conditions for single-cell separation, and inability to change the initial state of DNA mixing in test materials, etc. , to achieve the effects of weakening the effect of amplification bias, facilitating DNA map analysis, and increasing the probability of amplification

Pending Publication Date: 2020-09-29
INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, single cell separation technology depends on professional micromanipulation equipment and well-trained operators, and the conditions for single cell separation are limited
However, based on computer technology or manual splitting based on experienced experts, both rely on the acquisition of high-quality mixed maps,

Method used

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  • Method for performing STR typing on forensic mixed DNA sample
  • Method for performing STR typing on forensic mixed DNA sample
  • Method for performing STR typing on forensic mixed DNA sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1, detection standard substance mixed sample

[0051] 1. Preparation of mixed samples

[0052] The proportions of the 6 mixed samples are shown in Table 2.

[0053] Table 2

[0054]

[0055]

[0056] 2. Routine PCR amplification

[0057] use PCR Amplification Kit and operate according to the instructions.

[0058] The reaction system is 20 μL, consisting of 2 μL template, 2 μL Primer Set, 6μL Master Mix and 10 μL ddH 2 O composition.

[0059] The templates are the 6 mixed samples prepared in Step 1 respectively.

[0060] See the kit instructions for the reaction procedure, 29 cycles.

[0061] 3. Droplet digital PCR amplification

[0062] The reaction system is 20 μL, consisting of 2 μL template, 2 μL Primer Set, 6μL Master Mix and 10μL ddPCR Super Mix for probes (no dUTP).

[0063] The templates are the 6 mixed samples prepared in Step 1 respectively.

[0064] 1. Add 20 μL of the reaction system and 70 μL of droplet generating oi...

Embodiment 2

[0071] Embodiment 2, detect actual sample

[0072] Samples for sample: blood sample mixture (sample No. 12) and exfoliated cell mixture (sample No. 9, sample No. 10) from the center’s daily inspection.

[0073] 1. Extract the genomic DNA of each sample.

[0074] 2. Routine PCR amplification

[0075] Using AmpFLSTR TM Identifiler TM PCR Amplification Kit and operate according to the instructions.

[0076] In each reaction system, the content of genomic DNA was 0.5ng.

[0077] The templates are respectively the genomic DNA of each sample prepared in step 1.

[0078] 3. Droplet digital PCR amplification

[0079] The reaction system is 20 μL, consisting of 2 μL template (the DNA content in 2 μL template is 0.5 ng), 2 μL Primer Set, 6μL Master Mix and 10μL ddPCR Super Mix for probes (no dUTP).

[0080] The templates are respectively the genomic DNA of each sample prepared in step 1.

[0081] The method is the same as Step 3 of Example 1.

[0082] 4. Electrophoresis a...

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Abstract

The invention discloses a method for performing STR typing on a forensic mixed DNA sample. The invention provides a method for performing STR typing on a biological sample; and the method comprises the following steps of: extracting genome DNA of the biological sample, performing droplet type digital PCR amplification by taking the genome DNA as a template, sequencing an amplification product, andperforming STR typing according to a sequencing result, wherein the biological sample is a mixed biological sample. The invention also protects a method for performing STR typing on a DNA sample; andthe method comprises the following steps of: performing droplet type digital PCR amplification by taking the DNA sample as a template, sequencing an amplification product, and performing STR typing according to a sequencing result, wherein the DNA sample is a mixed DNA sample. The method provided by the invention has a huge application prospect in forensic DNA inspection.

Description

technical field [0001] The invention relates to a method for performing STR typing on forensic mixed DNA samples. Background technique [0002] With the improvement of the level of DNA testing, micro-contact test materials have gradually become the main test materials in DNA laboratories, and most of the samples obtained from such test materials are mixed DNA typing (such as the DNA of victims and suspects). admixture, admixture of two or more suspect samples, or admixture of a suspect with another unrelated individual, etc.). It is precisely these mixed samples that are of great practical significance in providing case clues, establishing the scope of investigation and providing court evidence. However, the DNA profile of the mixed sample is different due to the number of people mixed and the mixing ratio, resulting in many STR peak spectra, complex band patterns, and unstable peak patterns, resulting in difficult analysis of mixed sample results and high qualitative risks...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2531/113C12Q2563/159C12Q2525/151
Inventor 苑美青李万水凃政孟庆振李永久刘志芳王冲丰蕾徐秀兰
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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