Application and kit of reagents for detecting expression level of open reading frame 53 of chromosome 8
An open reading frame and expression level technology, applied in the biological field, can solve the problems that limit the wide application of breast cancer prognostic markers
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Embodiment 1
[0028] This example is used to illustrate the TCGA high-throughput data analysis of the expression changes of C8ORF53 in breast cancer.
[0029] 1. TCGA high-throughput data analysis process:
[0030] Log in to the TCGA portal UALCAN ( http: / / ualcan.path.uab.edu / index.html ) home page, click "Analysis", enter the gene name "C8ORF53", select TCGA dataset "Breast invasive carcinoma", search, click "Expression", and record the result. GraphPad 12.0 software was used to draw graphs, and the statistical method was T test, and P<0.05 was considered statistically significant.
[0031] 2. Results: The expression of C8ORF53 in breast cancer tissue was significantly higher than that in normal tissue (Pfigure 1 shown.
Embodiment 2
[0033] This example is used to illustrate the relationship between the high expression of C8ORF53 and the prognosis of breast cancer through TCGA high-throughput data analysis.
[0034] 1. TCGA high-throughput data analysis process:
[0035] Log in to the TCGA portal cBioPortal (http: / / www.cbioportal.org / ), select the tumor dataset "Breast Invasive Carcinoma (TCGA Provisional)", select "mRNA Expression z-Scores (RNA Seq V2RSEM)" for the omics data, and select the gene Enter "C8ORF53: EXP>=2", search, click "Survival" on the pop-up page, and record the result. Survival curves were drawn by Kaplan-Meier method, and the differences in survival curves were tested by log-rank test. P<0.05 was considered statistically significant.
[0036] 2. Results: High expression of C8ORF53 is not conducive to the overall survival of breast cancer patients (P=0.0127) ( figure 2 ).
Embodiment 3
[0038] This example is used to illustrate the preparation of reagents for detecting the expression of C8ORF53 and the preparation of kits for the prognosis of breast cancer patients (50 reactions).
[0039] 1. Trizol 50.0ml;
[0040] 2. Isopropanol 50.0ml;
[0041] 3. Chloroform 50.0ml;
[0042] 4. 50.0ml absolute ethanol;
[0043] 5. RNase-free water 5.0ml
[0044] 6. 1.0μM random primers (Random primers) 50.0μl;
[0045] 7. 2.0ml of 5×M-MLV buffer;
[0046] 8. 100.0μl of 10.0mM deoxynucleotide triphosphates (dNTPs);
[0047] 9. 40U / μl RNase inhibitor 50.0μl;
[0048] 10. 200U / μl M-MLV reverse transcriptase 50.0μl;
[0049] 11. ABI 2×PCR Mix 2.0ml;
[0050] 12. 10.0 μM C8ORF53 real-time fluorescent quantitative PCR specific primer 30.0 μl, the primer sequence is shown in Table 1;
[0051] 13. 30.0 μl of specific primers for 10.0 μM ACTB real-time fluorescence quantitative PCR. See Table 1 for the primer sequences.
[0052] Table 1 Fluorescence quantitative RT-PCR prim...
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