Application method of long non-coding RNA CRYM-AS1
An RNACRYM-AS1, long-chain non-coding technology, applied in the field of preparation of prognostic reagents for glioma patients, can solve the problems of low survival rate, high recurrence rate and unclear boundary of surrounding brain tissue in glioma patients, and achieve far-reaching clinical results. Meaning and Promotional Effects
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0025] Example 1 Preparation of LncRNACRYM-AS1 kit for the prognosis of glioma patients (50 reactions)
[0026] 1. RNA stabilization solution 50ml
[0027] 2. Isopropanol 100ml
[0028] 3. Chloroform 100ml
[0029] 4. Trizol 50ml
[0030] 5. Enzyme-free water 10ml
[0031] 6.1 μM random reverse transcription primer 50 μl
[0032] 7.5× reverse transcription buffer 200ml
[0033] 8. 10mM base triphosphate deoxynucleotides 100μl
[0034] 9.40U / μl RNase inhibitor 500μl
[0035] 10.200U / μl MMLV reverse transcriptase 50μl
[0036] 11. Premix ExTaq 50μl
[0037] 12. 10 μL ncRNACRYM-AS1 specific primer 30 μl
[0038] Forward primer 5'-CTGGAAGGCACGTAAGC-3',
[0039] Reverse primer 5'-GAGGCGGCGATGTATTG-3'.
[0040] 13. 10 μM U6snRNA specific primer 30 μl
[0041] The forward primer is 5'-ATTGGAACGATACAGAGAAGATT-3'
[0042] The reverse primer was 5'-GGAACGCTTCACGAATTTG-3'.
Embodiment 2
[0043] Example 2 Detection of LncRNACRYM-AS1 in Glioma Tissue
[0044] 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.
[0045]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 a liquid After the shape, transfer it to a tube, add 200μl / ml Trizol in chloroform to the Tube, shake it by hand for 15-30s, place it on ice for 5 minutes, and centrifuge at 12000g at 4°C for 15 minutes; carefully take the upper aqueous phase into a new tube, add pre-cooled iso Propanol 0.5ml / ml Trizol, mix well, put in refrigerator at -20°C for 20min, centrifuge at 12000g at 4°C for 10min; discard the supernatant, add 1-2ml ethanol diluted with 75% DEPC wa...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 