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Fluorescent dyes, specific amplification primer pairs and typing methods that can be used for nine-color fluorescent str typing

A technology for amplifying primers and fluorescent dyes, which is applied in the field of fluorescent dyes for nine-color fluorescent STR typing, can solve the problems of limiting the number of loci detected by the kit, and achieve good species specificity, sharp peak shape, and good system repeatability sexual effect

Active Publication Date: 2022-03-29
INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The most advanced DNA testing reagent, autosomes can simultaneously amplify and detect up to 27 loci, Y chromosome can reach 38, limited by factors such as the types of fluorescent dyes available and the design of primers, limiting the number of loci detected by the kit

Method used

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  • Fluorescent dyes, specific amplification primer pairs and typing methods that can be used for nine-color fluorescent str typing
  • Fluorescent dyes, specific amplification primer pairs and typing methods that can be used for nine-color fluorescent str typing
  • Fluorescent dyes, specific amplification primer pairs and typing methods that can be used for nine-color fluorescent str typing

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Effect test

Embodiment 1

[0029] Fluorescent dyes that can be used for nine-color fluorescent STR typing, the nine fluorescent dyes are FAM, TET, HEX, NED, ET598, ET618, ET635, ET650 and ET680. Among them, 8 fluorescent dyes FAM, TET, HEX, NED, ET598, ET618, ET635 and ET650 were used to fluorescently label the amplification primers of 41 autosomal STR loci and 1 sex-determining locus Amel; ET680 was used for labeling mark.

[0030] The structures of nine fluorescent dyes are shown in Table 1:

[0031] Table 1

[0032]

[0033]

[0034] The information of the 42 loci is shown in Table 2:

[0035] Table 2

[0036]

[0037]

Embodiment 2

[0039] The specific amplification primer pairs that can be used for nine-color fluorescent STR typing, and the specific amplification primers corresponding to 42 loci, are shown in Table 2, wherein PCRU represents the upstream primer, and PCRL represents the downstream primer.

[0040] table 3

[0041]

[0042]

[0043]

Embodiment 3

[0045] The nine-color fluorescent STR typing method of this embodiment is used to perform STR typing on the sample to be tested.

[0046] 1. Construction of the fluorescent system: 42 loci were labeled with 8 different fluorescent dyes, and ET680 was used to label the internal standard;

[0047] The eight different fluorescent dyes are: FAM, TET, HEX, NED, ET598, ET618, ET635, ET650 and ET680;

[0048] The 42 loci include 41 autosomal STR loci and 1 sex-determining locus Amel; the 41 autosomal STR loci are: D5S818, D21S11, D7S820, CSF1PO, D2S1338, D3S1358, vWA, D8S1179, D16S539 、D1S1677、PentaD、TPOX、TH01、 D22S1045、D18S51、FGA、D6S1043、D13S317、D12S391、PentaE、 D2S441、D1S1656、D19S433、D10S1248、D11S4463、D1S1627、D3S4529、 D1GATA113、D17S974、D6S1017、D4S2408、D9S2157、D6S474、D2S1776 , D18S853, D20S482, D14S1434, D20S1082, D17S1301, D12ATA63 and SE33;

[0049] The specific amplification primer pairs corresponding to the 42 loci are the nucleotide sequences of SEQ ID No.1 to SEQ ID No.84 in ...

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Abstract

The invention discloses fluorescent dyes, specific amplification primer pairs and typing methods that can be used for nine-color fluorescent STR typing, including nine kinds of fluorescent dyes, namely FAM, TET, HEX, NED, ET598, ET618, ET635, ET650 and ET680; 8 fluorescent dyes FAM, TET, HEX, NED, ET598, ET618, ET635 and ET650 are used to fluorescently label the amplification primers of 41 autosomal STR loci and 1 sex-determining locus Amel; ET680 is used for labeling mark. The typing method of the present invention has high sensitivity, stable results, complete and clear maps, and no obvious difference in repeated detection results; the stutter peak area ratio does not exceed 10%, the peak height ratio does not exceed 15%, and has good system repeatability. 42 genes Accurate typing results can be obtained in the detection of multiplex amplification system.

Description

technical field [0001] The invention relates to the technical field of fluorescent STR typing. Specifically, it is a fluorescent dye, a pair of specific amplification primers and a typing method that can be used for nine-color fluorescent STR typing. Background technique [0002] At present, commercially available DNA test kits are at most six-color fluorescent. For example, the applicant’s previous research such as Chinese patent document CN107254516 conducted a total of 25 loci including 24 autochromatic loci and 1 sex-determining locus. Fluorescent STR typing. [0003] The most advanced DNA testing reagents can simultaneously amplify and detect up to 27 loci for autosomes and 38 for Y chromosomes, which are limited by factors such as the types of fluorescent dyes available and the design of primers, which limit the number of loci detected by the kit. For trace degraded samples and identification of complex kinship, additional kit testing is required. Contents of the i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6876C09K11/06
CPCC09K11/06C12Q1/6876C12Q2600/156C09K2211/1088C09K2211/1044C09K2211/1048C09K2211/1096
Inventor 白雪叶健张涛宋文
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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