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

An application method and kit technology, applied in the fields of identification and DNA detection, can solve problems affecting trace DNA, etc., and achieve the effects of easy preservation, high DNA recovery rate, and good stability

Inactive Publication Date: 2009-04-01
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-100 20 (v / v), 1% NP-40 (v / v), 5% Chelex 100 (w / v) to prepare the sample collection solution. The specific preparation method is: prepare 1M Tris, adjust the pH value to 8.0 with HCl, use this 1M Tris-HCl, pH8.0 as the stock solution, dilute with water to prepare 30mM Tris-HCl, pH8.0, and other components according to the above concentration Add in sequence, and add proteinase K according to the volume of the collection solution used 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 into the eppendorf tube, take a sliver and put it into the wet, take out the sliver, wipe the part to be collected (such as sweat spots), and recover the sliver into the eppendorf tube .

[...

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 the sample collection solution. The specific preparation method is: prepare 1M Tris, adjust the pH value to 8.0 with HCl, use this 1M Tris-HCl, pH 8.0 as the storage solution, dilute with water to prepare 10mM Tris-HCl, pH 8.0, and other components in order according to the above concentrations Prepared, add proteinase K according to the specific volume of collection solution used before use to prepare a complete DNA sample collection reagent.

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

Embodiment 3

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

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

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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 identification in the fields of forensic evidence biology, anthropology, criminal investigation, etc., specifically a reagent for collecting and recovering trace 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 timely and effectively identifying criminals and fighting crime. [0003] In actual cases, suspects or persons responsible for incidents often leave latent fingerprints at the scene. In the past, on-site fingerprints have been compared based on morphological line features to achieve the purpose of individual identification. With the increasingly diverse and complex means of committing crimes, relying solely on morphological line features to identify indiv...

Claims

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Application Information

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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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