X chromosome MiniSTR fluorescent composite amplification reagent kit, preparation and use thereof

Abstract

The invention discloses an X chromosome parting kit and its preparation method and application. The present invention designs four pairs of public primer pairs, optimizedly select 12 X chromosome STRloci suitable for the genetic distribution of Chinese population and designs MiniSTR primers fro part of loci, and respectively adds the four pairs of public primers to the 5' end of the MiniSTR primers to obtain 24 additional primers; the use of the inventive kit can simultaneously amplify 12 X chromosome STR loci at a single-tube, combined with genetic markers of autosomal, Y chromosome and mitochondrial, the invention can effectively solve sister identification lack of parents, half-sister identification, atavism identification and the like special pro-prosecution case, at the same time canrealize the successful detection of DNA template as low as 0.05ng / 20mul. The inventive kit has advantages of high sensitivity, high detection rate of samples of a high degree of DNA degradation, lowpreparation cost and the like.

Description

technical field [0001] The invention relates to a PCR amplification kit, in particular to a fluorescent multiplex amplification kit for simultaneously detecting 12 X chromosome MiniSTR loci in a single tube. The present invention also relates to a preparation method of the fluorescent multiplex amplification kit and the The application of the kit in the field of judicial identification belongs to the field of X chromosome typing and identification. Background technique [0002] Short tandem repeat (short tandem repeat, STR) is a type of DNA sequence with length polymorphism formed by tandem repetition of 2 to 6 bases as the core unit in the human genome. The number of core units varies and the number of repetitions is different. The genetic polymorphisms that make up the STR. STR is widely distributed and numerous, accounting for about 10% of the human genome. The genetic information contained in it is unmatched by any polymorphic markers in the past. Therefore, STR is curr...

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Problems solved by technology

The only commercial kits are MentypeArgus X-8 (DXS8378, HPRTB, DXS7423, DXS7132, DXS10134, DXS10074, DXS10101 DXS10135) from Germany, which still has limitations in the number of loci and the ability to identify individuals in forensic DNA testing in my country

[0005] The number of loci that can be compounded and amplified by X-STR reported in domestic literature is small (usually 3-6), and most of them are detected by silver staining. When analyzing the amplification results, there must be time-consuming and low automation. Poor accuracy and other disadvantages, although the requirements for experimental equipment are not high, which meet the needs of the current domestic forensic basic-level laboratories, but cannot meet the higher requirements of most laboratories that already have automatic laser fluorescence genetic analyzers

Even if fluorescent detection is used, a number of primers used for amplification of single loci are directly added with fluorescent labels for multiple amplification, but with the increase of the number of primers in the amplification system, the interaction between primers is serious, and the reaction The kinetics become complicated, and the optimization of amplification conditions becomes extremely difficult, which becomes a difficulty in further increasing the number of multiple amplification loci

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