Application of incrnalnc_004208 and its detection reagents in the preparation of glioma prognosis reagents
A technology for detecting glioma and detection reagents, which is applied in the field of tumor molecular biology and can solve problems such as unexplained specific mechanisms
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Embodiment 1
[0036] Example 1 Transcriptome sequencing technology and lncRNA sequencing technology analysis of TMZ sensitive / resistant cell expression profiles
[0037] Transcriptome sequencing technology and lncRNA sequencing technology (Illumina HiSeq TM 2000), selected brain glioma T98G / U-118MG cell line and TMZ-resistant T98G-R / U-118MG-R cell line for expression profiling analysis. Before screening, we first used Cuffmerge software to merge the transcripts spliced from each sample, and removed transcripts with uncertain strand orientations to obtain the complete transcriptome information for this sequencing. The screening process is divided into the following 5 steps ( figure 1 A):
[0038] step1: Screening of the number of transcript exons: filter a large number of low-expression, low-confidence single-exon transcripts in the transcriptome splicing results, and select transcripts with exon numbers >= 2 (if there is no special requirement, the plant will Transcripts with exon numb...
Embodiment 2
[0049] Example 2 Screening and verification of differentially expressed lncRNAs in glioma TMZ-resistant cells
[0050] (1) Extraction of total cellular RNA (TRIzol method)
[0051] Cells were seeded in 6-well plates at 37°C, 5% CO 2 Incubate in the incubator. After the cells were completely attached to the wall, 1000 μl TRIzol was added to each well. After standing still until completely dissolved, transfer to a 1.5ml EP tube. Add 0.2ml of chloroform, shake vigorously for 15 seconds, and place at room temperature for 3 minutes. Centrifuge at 4°C and 12000rpm for 15min, and transfer the supernatant to a new EP tube. Add 0.5ml of isopropanol and let stand at room temperature for 10min. After centrifugation at 4°C and 12,000 rpm for 10 minutes, a gelatinous precipitate appeared at the bottom of the EP tube. Discard the supernatant, wash the RNA pellet with 1ml of 75℅ ethanol, and centrifuge at 4°C, 7500rpm for 5min, discard the supernatant again. After air-drying for 5 min...
Embodiment 3
[0066] Example 3 Correlation between patient survival and lncRNA expression
[0067] 104 patients with glioma were followed up by telephone, and they were asked in detail about the time of onset, grade, treatment, recurrence, other diseases, use of other drugs, recurrence and death time, etc. And registered the survival time and status. Survival analysis of the expression of LNC_004208 in brain glioma tissues and the survival time and status of patients found that the progression-free survival of patients with high expression ( Figure 4 A) and the average survival time was significantly shorter than that of patients with low expression or no expression ( Figure 4 B).
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