Composite amplification kit for simultaneously detecting autosomal and Y chromosome STR loci
A compound amplification and Y chromosome technology, which is applied in the field of compound amplification kits for simultaneous detection of autosome and Y chromosome STR loci, can solve the problems of increasing detection time, shortening detection time, detection failure, etc., and achieve strong detection materials Adaptability, shorter detection time, and accurate typing
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Embodiment 1
[0038] Example 1 Screening of autosomes and Y chromosome STR loci
[0039] 通过对DYS549、DYS522、DYS389I、DYS439、DYS447、DYS389II、DYS392、DYS576、DYS19、DYS388、DYS391、DYS635、DYS460、DYS456、DYS533、DYS520、DYS438、DYS527a / b、DYS449、DYS709、DYS622、DYS481、DYS437、 DYS390、DYS630、DYS448、Y_GATA_H4、DYS458、DYS607、DYS393、DYS552、DYS570、DYS385a / b、DYS593、DYS444、DYS643、Amelogenin、vWA、D10S1248、D2S441、D21S11、D18S51、D5S818、D7S820、D13S317、D16S539、FGA、 D2S1338, D22S1045, D1S1656, D19S433, D6S1043, D12S391, D8S1179, D3S1358, CSF1PO, Penta D, PentaE, TH01 and TPOX, a total of 62 loci, to ensure that the selected loci are compatible with commonly used kits (for example, ABI’s Identifiler and Yfiler Plus) loci have certain compatibility, and finally 20 Y chromosome STR loci were screened out (for example, locus names beginning with "DYS", including DYS391, DYS635, DYS576, DYS456, DYS385a / b, DYS438 , DYS448, DYS437, DYS19, DYS458, DYS533, DYS393, DYS389I / II, DYS439, DYS392, Y_GATA_H4, DYS390 and DYS481), 19 autosom...
Embodiment 2
[0044] Example 2 Group arrangement of 40 loci
[0045] DYS391, D3S1358, D13S317, D7S820, D16S539, Penta E, DYS635, DYS576 are the first group, and the fluorescent dye marker is 6-FAM; DYS456, TPOX, TH01, D2S1338, D6S1043, DYS385a / b, DYS438, DYS448 are the second group Group, fluorescent dye marker is HEX; DYS437, D19S433, vWA, DYS19, CSF1PO, D18S51, FGA is the third group, fluorescent dye marker is SUM; Amelogenin, D8S1179, D5S818, D21S11, D12S391, PentaD, DYS458, DYS533 are The fourth group, the fluorescent dye marker is LYN; DYS393, DYS389I / II, DYS439, DYS392, Y_GATA_H4, DYS390, DYS481 is the fifth group, the fluorescent dye marker is PUR, see the grouping diagram figure 1 shown.
Embodiment 3
[0046] Embodiment 3 specific primer design and the selection of primer concentration
[0047]With the increase of the number of primers in the multiplex amplification system, the mutual interference between primers at different loci becomes more and more serious, and the kinetics of the reaction system become more and more complex, so it is necessary to design a large number of primer sequences for complex testing And explore the specific concentration ratio between the primers in the compound amplification system, and finally ensure that more human autosomes and Y chromosome STR loci are compounded without reducing the specificity and sensitivity of the kit. Specifically, the following steps are included:
[0048] (1) Design of specific amplification primers
[0049] Before designing primers, it is necessary to analyze the properties of the target sequence to be tested, and select highly conserved regions with uniform base distribution for primer design.
[0050] a. Primer ...
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