Application of glioma prognostic marker Circ9:135881633|135883078
A glioma and prognosis technology, applied in the application of reagents in the preparation of glioma prognostic preparations, and the application field of circRNA markers, can solve the problems that the survival rate of glioma patients has not been significantly improved, and the survival rate is not optimistic.
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Embodiment 1
[0017] Example 1 Preparation of reagents for detecting expression of circular RNA Circ9:135881633|135883078 Kit for preparing prognosis of glioma patients (50 reactions)
[0018] 1. RNA stabilization solution 50ml
[0019] 2. Isopropanol 100ml
[0020] 3. Chloroform 100ml
[0021] 4. Trizol 50ml
[0022] 5. Enzyme-free water 10ml
[0023] 6. 1μM random reverse transcription primer 50μl
[0024] 7. 5× reverse transcription buffer 200ml
[0025] 8. 100μl of 10mM base triphosphate deoxyribonucleotides
[0026] 9. 40U / μl RNase inhibitor 500μl
[0027] 10. 200U / μl MMLV reverse transcriptase 50μl
[0028] 11.Premix Ex Taq 50μl
[0029] 12. 10μM Circ9:135881633|135883078 real-time fluorescence quantitative PCR specific primer 30μl
[0030] Circular RNA Circ9:135881633|135883078 Forward primer:
[0031] 5'-AATGGTGGATGCCCTGAT-3',
[0032] Circular RNA Circ9:135881633|135883078 reverse primer:
[0033] 5'-TGTGCTCCTGCTCATACTGG-3';
[0034] 13. 10μM GAPDH specific primer 30μl...
Embodiment 2
[0037] Example 2 Detection of expression level of circular RNA Circ9:135881633|135883078 in brain tissue samples
[0038] 1. Collect the glioma tissue to be tested, put it into a cryopreservation tube filled with RNA stabilization solution, and put it in a -80°C refrigerator for later use.
[0039] 2. Extraction of RNA in the tissue: take 0.1g of the above-mentioned tumor and normal brain tissue, rinse with an appropriate amount of normal saline, and put it into a mortar (the mortar must be wrapped with tinfoil in advance and baked in an oven at 180 degrees for 6-8 hours). hours), add an appropriate amount of liquid nitrogen to grind the tissue to powder, add 0.6ml Trizol (MRC company) and continue grinding for a few minutes, add 0.4ml Trizol again, transfer the mixture in the mortar to a 1.5ml enzyme-free Tube tube, and put it on ice Lyse for 15 minutes. After the lysis, transfer to a 1.5ml enzyme-free tube, centrifuge at 12000rpm at 4°C for 10min, and transfer the supernata...
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