Noninvasive antepartum fetal alpha<-SEA> type thalassemia gene mutation detection library building method, detection method and kit
A library construction, -SEA technology, applied in chemical libraries, biochemical equipment and methods, DNA preparation, etc., can solve the problems of lack of homozygous type, inability to further determine the presence or absence of maternal mutations, typing errors, etc. achieve the effect of improving specificity
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[0090] 1. Targeting α-thalassemia gene-α SEA Design synthetic primers for mutation sites:
[0091] The upstream primer and downstream primer are located on the left and right of the detection point, respectively. The 3' end of specific primer 1 and the 5' end of specific primer 2 on the same side overlap by 10-15 bases. The 5' end of specific primer 1 was modified with biotin. The 5' end of specific primer 2 contains adapter sequences for high-throughput sequencing libraries. Specifically, the primer sequences are shown in Table 1.
[0092] Table 1
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[0097] 2. Linker sequence with specific tag sequence and sample tag sequence (index):
[0098] ADT-F:
[0099] CAAGCAGAAGACGGCATACGAGATNNNNNNNATTAAGGGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT (SEQ ID NO: 45);
[0100] ADT-R: pGATCGGAAGAGC (SEQ ID NO: 46).
[0101] Among them, NNNNNNNNN is a specific tag sequence, ATTAAGG is a sample tag sequence (index), and the above linker sequences ...
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