Y chromosome MiniSTR typing reagent kit, preparation and use thereof

Abstract

The invention discloses a Y chromosome parting kit and its preparation method and application. The present invention designs three pairs of public primer pairs, optimizedly select 12 X chromosome STR loci suitable for the genetic distribution of Chinese population and designs MiniSTR primers fro part of loci, and respectively adds the three 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, avoids the competition between these primers and the formation of heterozygosity dimers, increases the amplification efficiency of each locus, wherein, the length of amplification products can be less than the 280bp, and has a high sensitivity (as low as 30pg), a higher STR parting success rate can be obtained and the amplification efficiency of medical examiner DNA degradation samples can be improved. The inventive kit has advantages of high sensitivity, high specificity, high detection efficiency, low preparation 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 Y chromosome MiniSTR loci in a single tube, and also relates to a preparation method of the fluorescent multiplex amplification kit and the The application of the kit in forensic identification belongs to the field of 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 currently the most widely used genetic marker fo...

AI Technical Summary

Problems solved by technology

[0008] There are still the following problems in its application in domestic forensic medicine: ①Compared with commercial autosomal STR kits, the genetic distribution of Y-STR has more significant regional, racial, and ethnic differences, and the imported kits are based on foreign Caucasian populations. Development, most loci have poor gene frequency distribution and low personal identification ability in the Chinese population, such as DYS389I, DYS391, DYS393, DYS437, DYS438, etc., to a certain extent, it is difficult to meet the work needs of forensic DNA testing; ②Reagent prices Expensive, which not only consumes a lot of national funds for handling cases, but also limits the domestic application range of DNA technology

However, since the amplified fragments of this kit and the above-mentioned two imported Y-STR commercial kits are all distributed between 100-400bp, it is difficult to detect highly degraded DNA samples such as old bones, teeth, and decayed tissues, as well as trace amounts. For low-content DNA samples such as bloodstains, fingernails, and hair, large fragments of Y-STR loci cannot be effectively amplified, and the amplification efficiency decreases significantly with the increase of fragments, alleles are missing or completely lost etc.

In addition, the allele frequency distribution of DYS434, DYS438, DYS439, DYS453, DYS513, Y-GATA-A10, and Y-GATA-A8 loci contained in the kit is poor, the individual recognition rate is low, and the detection efficiency of the whole system Still needs to be further improved

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