Glioma prognostic marker circ15:98707562|98708107 and applications
A glioma and prognosis technology, applied in the application of circRNA markers, and the application field of reagents in the preparation of glioma prognosis preparations, can solve the problems of the survival rate of glioma patients is not significantly improved, and the survival rate is not optimistic.
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Embodiment 1
[0018] Example 1 Preparation of reagents for detecting the expression of circRNA circ15:98707562|98708107 for the preparation of a kit for the prognosis of glioma patients (50 reactions)
[0019] 1. RNA stabilization solution 50ml
[0020] 2. Isopropanol 100ml
[0021] 3. Chloroform 100ml
[0022] 4. Trizol (from Molecular Research Center company) 50ml
[0023] 5. Enzyme-free water 10ml
[0024] 6. 1μM random reverse transcription primer (Thermo Company) 50μl
[0025] 7. 5× reverse transcription buffer (Thermo company) 200ml
[0026] 8. 10mM base triphosphate deoxynucleotide (Thermo company) 100μl
[0027] 9. 40U / μl RNase inhibitor (Thermo Company) 500μl
[0028] 10. 200U / μl MMLV reverse transcriptase (Thermo Company) 50μl
[0029] 11. Premix Ex Taq (Thermo Company) 50μl
[0030] 12. 10μM circRNA circ15:98707562|98708107 real-time fluorescence quantitative PCR specific primer 30μl
[0031] circRNA circ15:98707562|98708107 Forward primer: 5'-CCGATGTGTGAGAAGACCA-3',
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Embodiment 2c
[0036] Example 2 The relationship between the expression level of circRNA circ15:98707562|98708107 in glioma tissue and prognosis
[0037] 1. Preservation of glioma tissue: collect the glioma tissue to be tested and store it in a cryopreservation tube filled with RNA stabilization solution, and put it in a -80°C refrigerator for later use.
[0038] 2. Extraction of RNA in tissues: Take an appropriate amount of specimen, add liquid nitrogen to the mortar after baking at 180°C for 6-8 hours, grind the specimen, grind to powder, add 1ml Trizol mortar specimen to the mortar, and grind into After the liquid was transferred to a tube tube, it was lysed on ice for 15 minutes. After the lysis, centrifuge at 12000rpm for 10min at 4°C, and transfer the supernatant to a new tube. Add 200μl of chloroform to the tube, shake it by hand for 15-30s, place on ice for 15min, centrifuge at 12000rPm at 4°C for 15min; carefully take the upper aqueous phase into a new tube, add 0.5ml of pre-cooled...
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