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Fluorescently-labeled X-InDel locus composite amplification system and application thereof

A compound amplification system and fluorescent labeling technology, which can be used in fluorescence/phosphorescence, microbial determination/inspection, biochemical equipment and methods, etc. Technically fast, highly sensitive results

Active Publication Date: 2014-11-05
ACADEMY OF FORENSIC SCIENCE
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  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

There have been some international publications on the emerging genetic marker InDel on the X chromosome, but domestic related research is still relatively lacking, and even the data on the Asian population is relatively scarce

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1 Application of X-InDel Site Fluorescence Multiplex Amplification System to Carry out Parent-child Identification and Individual Identification

[0074] Steps:

[0075] 1. DNA extraction

[0076] Genomic DNA extraction by Chelex-100 method: add 1mL double distilled water to a 1.5mL centrifuge tube, then add 3μL whole blood or 3×3mm blood spot, mix carefully; keep warm at room temperature for 30min, shake intermittently; centrifuge at 14,000r / min for 5min; Carefully remove the supernatant. Add 200 μL of 10% Chelex100 (100 mesh) to the precipitate; incubate at 56°C for 30 minutes; shake at high speed for 5-10 sec; bath in boiling water for 8 min; centrifuge at 14,000 r / min for 5 min, and take the supernatant for PCR amplification.

[0077] 2. PCR amplification

[0078] The total reaction volume was 15.0 μL, including 7.5 μL of PCR reaction buffer (Buffer), 3.0 μL of primer mix (Primer pair Mix), 0.5 μL of DNA polymerase, 3.0 μL of template DNA, and 1 μL of ult...

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PUM

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Abstract

The invention provides a fluorescently-labeled X-InDel locus composite amplification system. By means of the system, eighteen insertion-deletion genetic polymorphism loci on an X chromosome can be compositely amplified, wherein the eighteen loci are respectively labeled with three fluoresceins FAM, HEX and TAMRA. The composite amplification system is high in sensitivity and individual identification rate, is high in polymorphism, is good in stability and repeatability, is accurate in typing results and can satisfy a practical requirement. A kit can be manufactured on the basis of the composite amplification system, can be used for paternity test, dyad paternity test, grandparent and grandchild test, sibling test and individual identification. New technologies can be provided for the fields such as anthropology, medical genetics and the like.

Description

technical field [0001] The present invention relates to a detection of polymorphic genetic markers in the human genome, in particular to a fluorescent marker complex amplification system and its application. Background technique [0002] STR (Short Tandem Repeats, STR) typing technology is a commonly used technique in forensic biology research and identification. For now, STR typing is considered to be the most standard and authoritative approach and method in forensic DNA identification, which can solve most practical problems. However, with the widespread application of STR loci in forensic DNA analysis, its defects have attracted increasing attention from the academic community. For example, the high mutation rate of STR loci is not conducive to the interpretation of the results of paternity testing; the long PCR amplicon makes it difficult to realize DNA typing of degraded samples; the limited number of STR loci is not conducive to the identification of complex kinship...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2563/107C12Q2531/113
Inventor 李成涛孙宽赵书民张素华
Owner ACADEMY OF FORENSIC SCIENCE
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