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Reagent and kit for acquisition and recovery trace amount DNA and use method

A reagent and trace technology, which is applied in the field of DNA detection and identification, can solve the problems affecting trace DNA, etc., and achieve the effect of easy preservation, simple operation and good stability

Inactive Publication Date: 2012-05-30
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The limitation of on-site DNA sample collection technology in the early stage has greatly affected the detection efficiency of trace DNA, especially sweat spot fingerprint DNA

Method used

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  • Reagent and kit for acquisition and recovery trace amount DNA and use method
  • Reagent and kit for acquisition and recovery trace amount DNA and use method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Press 30mM Tris-HCl, pH8.0, 50mM NaCl, 2mM MgCl 2 , 1% TritonX-10020 (v / v), 1% NP-40 (v / v), 5% Chelex 100 (w / v) to prepare sample collection liquid. The specific preparation method is: prepare 1M Tris, adjust the pH value to 8.0 with HCl, use 1M Tris-HCl, pH 8.0 as the storage solution, dilute with water to prepare 30 mM Tris-HCl, pH 8.0, and other components according to the above concentration Prepare them one by one, and add proteinase K according to the specific volume of the collection solution before use to prepare a complete DNA sample collection reagent.

[0050] Use this DNA sample collection solution to collect 50 sweat samples, and collect DNA according to the following steps:

[0051] 1. According to the size of the collection area, take 10ul-20ul sample collection solution to the eppendorf tube, take a cotton sliver and put it in the eppendorf tube, take out the cotton sliver, wipe the part to be collected (such as sweat spots), and collect the sliver into the e...

Embodiment 2

[0057] Press 10mM Tris-HCl, pH8.0, 10mM KCl, 0.5mM MgCl 2 , 0.2% Tween 20 (v / v), 0.2% NP-40 (v / v), 5% Chelex 100 (w / v) to prepare sample collection liquid. The specific preparation method is: prepare 1M Tris, adjust the pH value to 8.0 with HCl, use 1M Tris-HCl, pH 8.0 as the storage solution, dilute with water to prepare 10mM Tris-HCl, pH 8.0, and prepare the other components in sequence according to the above concentrations Before use, add proteinase K according to the specific collection fluid volume to prepare a complete DNA sample collection reagent.

[0058] Use this DNA sample collection solution to collect another 50 sweat samples, collect DNA according to the steps in Example 1, and then use the 10ul amplification system of the Profiler Plus (ABI Company) kit to carry out STR-DNA amplification, and successfully carry out STR Subtype.

Embodiment 3

[0060] In the murder case, gloves of unknown origin were found at the scene. Press 30mM Tris-HCl, pH 8.0, 50mM KCl, 2mMMgCl 2 , 1% Tween 20 (v / v), 1% NP-40 (v / v), 5% Chelex 100 (w / v) to prepare a sample collection solution. The specific preparation method is as follows: prepare 1M Tris, adjust the pH value to 8.0 with HCl, use 1M Tris-HCl, pH 8.0 as the storage solution, dilute with water to prepare 30mM Tris-HCl, pH 8.0, and prepare the other components according to the above concentrations. Before use, add proteinase K according to the specific collection fluid volume to prepare a complete DNA sample collection reagent.

[0061] The DNA sample collection solution was used to sample different parts of the glove. After DNA detection and comparison, it was found that the DNA was different from the victim's DNA (Figure 1, left). The DNA of the suspect was tested, and it was found to be completely consistent with the residual DNA detected in the gloves (Figure 1, right). At the sa...

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Abstract

The invention relates to an agent formula used for collecting and recovering trace amount of DNA by a one-step method and a corresponding kit. The agent formula is mainly characterized in that the agent which is used for collecting and recovering trace amount of DNA and consists of a pH buffer system, salt ions, a surfactant, chelate resin and prolease is adopted, can furthest recover spot forensic material evidence DNA and has good effect on the recovery of sweat fingerprint DNA residual on different materials. The collected DNA can be directly used for the subsequent analysis without the steps of DNA extraction and purification. Compared with the traditional methods, the method has the characteristics of high DNA recovery ratio, simple and convenient operation, low cost, high accuracy and good repeatability, and the developed kit based on the formula can be directly used for collecting DNA samples of the crime scene in the first time.

Description

Technical field [0001] The invention relates to DNA detection and identification technologies such as individual identification and paternity testing in the fields of forensic evidence biology, anthropology, criminal investigation, etc., and specifically relates to a reagent for collecting and recovering trace amounts of DNA and a kit containing the reagent and Application method. Background technique [0002] In the work of forensic investigation, the identification and detection of forensic evidence at the crime scene plays a vital role in identifying criminals in a timely and effective manner and fighting crimes. [0003] In actual cases, the suspect or the person responsible for the incident often left sweat latent fingerprints at the scene. In the past, on-site fingerprints have been compared from morphological line features to achieve the purpose of individual identification. With the increasing diversification and complexity of crime methods, the identification of individua...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 罗瑛柳海涛唐文如李安史斌
Owner KUNMING UNIV OF SCI & TECH
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