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Fluorescence multiplex amplification system, kit and application of 36 y-chromosomal str loci and one y-indel
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A compound amplification system and y-indel technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problem of cumulative individual identification and cumulative non-parent exclusion rate to be further improved, etc. question
Active Publication Date: 2021-08-31
SUZHOU MICROREAD GENETICS
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At present, the existing kits on the market have a small number of loci, and there is no multiple amplification system that includes these 35 loci at the same time. The cumulative individual recognition ability and cumulative non-parent exclusion rate still need to be further improved
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Embodiment 1
[0043] Example 1 36 Y selected STR loci and a Y-Indelfluorescence labeled primer design and composition embodiments
[0044] The product in the selected locus, the fusion MPS 20 and core 15 preferably loci loci, while adding a Y-Indel site, further comprising a site-specific DYF404S1.
[0045] The loci 36 and a Y-Indel divided into five groups, through experiments repeated several times to determine the optimal combination of fluorescent dyes embodiment, Y-Indel, DYS393, DYS570, DYS19, DYS392, DYS549, Y-GATA-H4 , DYS444 labeled with FAM; DYS460, DYS458, DYS481, DYS635, DYS448, DYS533, DYS596 with HEX-labeled; DYS456, DYS389I, DYS390, DYS389II, DYS438, DYS447 and DYS596 with L552 marker; DYS391, DYS439, DYS437, DYS385a / b, DYS643, DYS518 labeled with LR600; DYS576, DYF404S1, DYF387S1, DYS627, DYS527a / b, DYS557 labeled with TET-592.
[0046] A pair of primers designed for each of these loci, primer pairs amplified single experiment, ensure that does not cause non-specific amplif...
[0058] Before testing, the product was denatured and mixed with internal standard molecular weight, the product IuI + 0.5ul + 8.5ul of standard molecular weight in the HIDI (deionized formamide), centrifuged briefly mixing, immediately after 5min denaturation 95 deg.] C ice bath 10min, electrophoresis detected by ABI series genetic analyzer, such as figure 1 . As can be seen from the figure, each locus can be accurately typing, better specificity, nonspecific band does not appear, the peak height of each locus balance is also good.
Embodiment 3
[0060] Species-specific experiments
[0061] Take pigs, cattle, sheep, chickens template for amplifying, detecting Y chromosome 36 STR loci and a Y-Indel species-specific multiplex amplification system, as a result figure 2 Indicated. By amplification of the other species, not amplified bands, 34 pairs of primers described species of genus-specific preferred.
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Abstract
The invention provides a fluorescencemultiplex amplification system, kit and application of 36 Y-chromosomal STR loci and a Y-Indel, belonging to the technical field of biological detection. The kit can simultaneously amplify and analyze the STR loci DYS393 and DYS570 、DYS19、DYS392、DYS549、Y‑GATA‑H4、DYS391、DYS439、DYS481、DYS635、DYS448、DYS533、DYS456、DYS389I / II、DYS390、DYS438、DYS576、DYS460、DYS458、DYS437、DYS385a / b、DYS643、DYF387S1 a / b, DYS627, DYS449, DYS518, DYS444, DYS447, DYS596, DYF404, DYS527, DYS557 and Y-Indel sites, the present invention uses six-color fluorescent markers, the size of the amplified product is between 70-490bp, and the operation is simple, And provided a high cumulative individual discrimination and cumulative non-parent exclusion rate.
Description
Technical field [0001] The present invention belongs to the field of biological detection, and is used to detect short series repeating sequence locus polymorphisms in the human genome. Specifically, the present invention relates to 36 Y-chromosome Str locations and 1 Y-INDEL fluorescent composite amplification system. , Kit and its application. Background technique [0002] Human Y chromosome specific short series repeating sequences are Y-STR (SHORT TANDEM REPEAT, STR), which is unique to human men, in accordance with the parent haplotype, in legologic parent-child identification and individual identification, human population, and evolution There is a unique application value in the study, and there is also an important application prospect in research on non-invasive post-prenatal gene diagnosis. Y-STR inspection technique is a supplementary means of testing of common-stained STR test techniques. This technology has a very important application value for the suspects to inves...
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