Glioma prognostic marker circ1:246186743|246186942 and its application
A glioma and prognosis technology, applied in biochemical equipment and methods, microbial determination/examination, DNA/RNA fragments, etc., can solve the problems of unsatisfactory curative effect and easy recurrence.
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Embodiment 1
[0016] Example 1: Preparation of circRNA circ1:246186743|246186942 kit for the prognosis of glioma patients (50 reactions)
[0017] 1. RNA stabilization solution 50ml
[0018] 2. Isopropanol 100ml
[0019] 3. Chloroform 100ml
[0020] 4. Trizol 50ml
[0021] 5. Enzyme-free water 10ml
[0022] 6.1 μM random reverse transcription primer 50 μl
[0023] 7.5× reverse transcription buffer 200ml
[0024] 8. 10mM base triphosphate deoxynucleotides 100μl
[0025] 9.40U / μl RNase inhibitor 500μl
[0026] 10.200U / μl MMLV reverse transcriptase 50μl
[0027] 11.Premix Ex Taq 50μl
[0028] 12.10μM circRNA circ1:246186743|246186942 real-time fluorescence quantitative PCR specific primer 30μl
[0029] Forward primer: 5'-CACGACTTGGCTGAGGAA-3',
[0030] Reverse primer: 5'-CCCCACATCACAGGCACA-3'.
[0031] 13.10 μM GAPDH specific primer 30 μl
[0032] The forward primer is 5′-ATCATCAGCAATGCCTCCT-3′,
[0033] The reverse primer was 5'-CATCACGCCACAGTTTCC-3'.
Embodiment 2
[0034] Example 2: The relationship between the expression level of circRNA circ1:246186743|246186942 in glioma tissue and prognosis
[0035] 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.
[0036] 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 it to powder, add 1ml Trizol mortar specimen in the mortar, and grind it into After the liquid was transferred to a 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 200 μl of chloroform to the tube, shake it by hand for 15-30 seconds, place it on ice for 5 minutes, centrifuge at 12000 rpm for 15 minutes at 4°C; carefully take the upper layer of the aqueous phase into a new tube, add 0.5ml of pre-coo...
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