Biomarker for early breast cancer diagnosis
A breast cancer, drug technology, applied in the field of biomedicine
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Embodiment 1
[0041] Example 1 Screening for Gene Markers Related to Early Breast Cancer
[0042] 1. Sample collection
[0043] 30 breast cancer tissue samples and corresponding paracancerous tissue samples were collected from 30 cases of stage I-II, and 5 cases were randomly selected for high-throughput sequencing. All cases did not receive chemotherapy and radiotherapy before surgery to exclude other tumors. Diseases, autoimmune diseases, and severe chronic diseases, all patients gave informed consent and were approved by the organization's ethics committee.
[0044] 2. Preparation of RNA samples
[0045] Cut the tissue into pieces with scissors, add 1ml Trizol, shake on the shaker for 1min; place at room temperature for 10min to completely decompose the ribosomes; then add 200μl chloroform (chloroform), close the tube cap tightly, shake vigorously for 15s, and let stand at room temperature for 10min Finally, centrifuge at 11000rpm for 15min at 4°C; transfer the water sample layer to a ...
Embodiment 2
[0056] Example 2 QPCR sequencing verification of differential expression of LINC01929 gene
[0057] 1. Using the previously collected tissue samples of 30 cases of stage I-II breast cancer and paracancerous tissue samples to conduct large-scale QPCR verification of LINC01929.
[0058] 2. The RNA extraction operation steps are the same as in Example 1
[0059] 3. QPCR
[0060] 1) Reverse transcription reaction
[0061] The lncRNA reverse transcription was carried out using the FastQμant cDNA First Strand Synthesis Kit (Product No.: KR106) from Tiangen Company.
[0062] First remove the genomic DNA reaction, add 5×gDNA Bμffer 2.0μl in the test tube, total RNA 1μg, add RNase Free ddH 2 O Make the total volume to 10μl, heat in a water bath at 42°C for 3min, then add 10×Fast RT Bμffer 2.0μl, RT Enzyme Mix 1.0μl, FQ-RT Primer Mix 2.0μl, RNase Free ddH 2 O 5.0 μl, after mixing, add to the above test tube and mix together a total of 20 μl, heat in a water bath at 42°C for 15min, t...
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