DNA (Deoxyribonucleic Nucleic Acid) probe library for hybridization with BCR or ABL gene and method for enriching BCR-ABL gene segments
A technology of DNA probes and gene fragments, applied in the field of enrichment and extraction of BCR-ABL gene fragments
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Embodiment 1
[0151] Example 1: Enrichment and detection of BCR-ABL fusion genes in K-562 cell line and MDA-MB-231 cell line
[0152] 1. Prepare the DNA sample bank of the cell line to be tested separately
[0153] 1. Extract K-562 cell line (human chronic leukemia cell line, from ATCC cell bank, catalog number: ATCC-CCL-243) and MDA-MB-231 cell line (human breast cancer cell line, from ATCC cell bank, catalog number: HTB-26), and then fragment it (the DNA sample bank obtained in this way is called "DNA sample bank derived from the whole genome")
[0154] 1.1 DNA extraction
[0155] Using Qiagen Blood&Tissue DNeasy Kit (Product No.: 69506), the whole genome DNA was extracted from the two cell lines respectively. Operate according to the instructions in the manual.
[0156] Use spectrophotometer and gel electrophoresis system to detect the quality and concentration of DNA. The 260nm absorbance of dsDNA is greater than 0.05, and the absorbance A260 / A280 ratio between 1.8 and 2 is qualif...
Embodiment 2
[0233] Example 2: Enrichment and detection of BCR-ABL fusion gene in KU812 cell line
[0234] Using the probe library consisting of SEQ ID NO.1 to SEQ ID NO.31 obtained in Example 1, the same method as in Example 1 was used to detect the KU812 cell line (human chronic leukemia cell line, from the ATCC cell bank, Item No.: ATCC-CRL-210) BCR-ABL gene, BCR:ABL gene fusion was found in KU812 cell line, the fusion points are Chr22:23632851, Chr9:133650197.
[0235] After DNA extraction, PCR amplification and Sanger sequencing, the above mutations have been verified.
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