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Primer design method for multiple microRNAs multiplex detection

A technology of primer sequences and substances, applied in the field of forensic medicine, can solve the problems of difficult to meet the requirements of rapid detection of miRNAs labeling at the same time

Active Publication Date: 2016-05-11
SICHUAN UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Real-time quantitative PCR technology and gene chip technology are difficult to meet the requirements of simultaneous and rapid detection of multiple miRNAs markers for forensic trace biological samples

Method used

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  • Primer design method for multiple microRNAs multiplex detection
  • Primer design method for multiple microRNAs multiplex detection
  • Primer design method for multiple microRNAs multiplex detection

Examples

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Effect test

Embodiment 1

[0061] Example 1 Using the primers designed by the present invention to detect 10 irrelevant Han individual bloodstains or menstrual blood spots or semen spots

[0062] In this example, four reported miRNAs were selected: miR214, miR451a, miR888 and miR891a. The information of the 4 miRNAs loci is shown in Table 3.

[0063] Table 34 miRNA loci

[0064]

[0065] According to the primer design method of the present invention, specific reverse transcription primers (REP) of 4 miRNAs were obtained, and the nucleotide sequences of each primer are shown in Table 4.

[0066] Table 4 specific reverse transcription primers

[0067]

[0068] In the sequences described in Table 4, before the "-" symbol is the ADP sequence, and after the "-" symbol is the specific sequence P1 complementary to the 3' end of the target miRNA marker.

[0069] According to the primer design of the present invention, 4 miRNAs-specific PCR primers PF and 1 common PCR primer PR were obtained, and the nu...

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Abstract

The invention belongs to the technical field of forensic medicine, and particularly relates to a method for designing primers for compositely detecting multiple microRNAs (Micro Ribonucleic Acids), aiming at the technical problem of determining the tissue source of a body fluid blackspot by detecting microRNAs. According to the technical scheme, the primers for detecting the microRNAs include a specific reverse transcription primer REP, a forward primer PF and a reverse primer PR, wherein the specific reverse transcription primer REP sequentially comprises joint sequences ADP and P1 from a 5' terminal to a 3' terminal; the ADP sequentially includes a special sequence NHS and a TTTTC sequence of a nonhuman genome from the 5' end to the 3' end; the P1 refers to 6-8 basic groups which are complementary with the 3' terminal of an miRNA marker to be detected. The primers disclosed by the invention can be used for establishing the co-platform detection of miRNA and forensic medicine DNA genetic markers, can be used for compositely amplifying a plurality of miRNAs, can obtain more tissue source information from common forensic medicine trace detection material and has outstanding application value in the field of forensic genetics.

Description

technical field [0001] The invention belongs to the technical field of forensic medicine, and in particular relates to a primer design method for compound detection of multiple microRNAs. Background technique [0002] Forensic evidence identification can obtain useful information for case investigation through the detection of biological samples left at the crime scene. Forensic biological examination materials are different from general biological examination materials. Most of the crime scenes are some bodily fluid spots, such as blood stains, saliva spots, and semen spots. Determining the tissue source of these bodily fluid spots is crucial to clarifying the nature of the case. For example, in a rape case, proving that the bodily fluid spots on the suspect's body come from the victim's vaginal secretions or menstrual blood will make it difficult for the suspect to dispel it. Interpreted as formed by everyday contact or physical aggression. Another example is in child se...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/10
CPCC12Q1/6811C12Q2525/207C12Q2531/113C12Q2565/125
Inventor 侯一平颜静李岩魏蔚张霁
Owner SICHUAN UNIV
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