Human chromosome 10 centromere probe and preparation method and kit thereof

[0035] The nucleotide sequence of the human chromosome 10 centromere probe of this embodiment is shown in SEQ ID NO.1.

[0036] Two, the specific embodiment of the preparation method of human chromosome 10 centromere probe of the present invention

[0038] This embodiment illustrates the preparation process of the human chromosome 10 centromere probe in Example 1, which is prepared by a nested PCR method, including the following steps:

[0039] 1) Using human genomic DNA as a template, the first round of PCR reaction was performed using primers Chr10-1-F and Chr10-1-R;

[0040] The primer sequences used are as follows:

[0041] Chr10-1-F: 5'-CGGGATTTTTTCATATAACGCTA-3';

[0042] Chr10-1-R: 5'-GTATTTCCAAACTGCTCGACT-3';

[0043] 2) Then, the first-round PCR product was diluted 100 times as a template, and the primers Chr10-2-F and Chr10-2-R were used for the second round of PCR reaction to obtain the product;

[0044] The primer sequences used are as follows:

[0045] Chr10-2-F: 5'-TTCTCAGTAACTACTTTGTG-3';

[0046] Chr10-2-R: 5'-TTTGTCTAGTTTTTAATAC-3'.

[0047] Both the first and the second round of PCR reaction procedures adopt the Touchdown PCR method.

[0048] The nucleotide sequences of primers Chr10-1-F and Chr10-...

[0055] The human chromosome 10 centromere probe kit in this embodiment is the working solution containing the probe in Example 1.

[0056] The working solution is made of human chromosome 10 centromere probe diluent, 1×TE buffer, Cot-1 DNA blocking agent, and hybridization buffer at a volume ratio of 1:1:1:7. Human chromosome 10 centromere probe diluent was prepared by diluting the nested PCR product of Example 2 with 1×TE buffer solution (10mM Tris-HCl, 1mM EDTA pH 8.0) at 1:15, and the concentration was 5ng / μL. Specifically, 1 μL human chromosome 10 centromere probe diluent, 1 μL 1×TE buffer buffer, 1 μL Cot-1 DNA blocking agent, 7 μL hybridization buffer, and the liquid components were thoroughly mixed.

[0057] 4. Experimental example

[0058] Taking human peripheral blood lymphocytes as samples, the kit of Example 3 was used to detect the specificity and brightness of the fluorescent signal of the centromere probe of chromosome 10 by using the rapid fluorescence in sit...