Detection kit for fusion mutation of ROS1 and various genes
A detection kit, fusion mutation technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc. The effect of wide clinical application, wide detection range and fast detection speed
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0125] Using the system of the present invention to detect plasmids, the experimental plasmid template (containing CD47-E6 and ROS1-E32 gene fusion), and the method for detecting ROS1 gene fusion mutation using the above-mentioned fluorescent PCR are as follows:
[0126] (1) Plasmid processing and extraction
[0127] The plasmid was extracted using TIANGEN (HighPure Plasmid kid, DP116) plasmid extraction kit, and the specific extraction steps were performed according to the kit instructions. The extracted DNA is dissolved in Tris-HCl (10mM, pH 8.0), and the extraction quality is detected by UV spectrophotometer to determine its concentration, and then the DNA concentration is adjusted to 20ng / ul with Tris-HCl (10mM, pH 8.0) solution as the dilution mother liquid .
[0128] According to formula C Copy concentration =(C Concentration X6.02X10 23 ) / MW DNA Dilute the wild-type plasmid to 10 6 Copies / μl.
[0129] (2) Establish a PCR amplification reaction system:
[0130] Use the cDNA ...
Embodiment 2
[0147] Using the present invention to detect clinical samples, 115 cases of clinical lung cancer paraffin-embedded tissue samples to be tested by our company, 69 cases of males and 46 cases of females, aged 35-76 years old, average age 56 years old, median age 52 years old . The steps for detecting ROS1 gene fusion mutations of 115 clinical samples by using the specific primers and probe fluorescent PCR system of the present invention are as follows:
[0148] (1) Sample processing and RNA extraction
[0149] a. Take about 1×1cm2 of each lung cancer sample slice (about 4-5 slices in total) and place it in a centrifuge tube (self-prepared), add 1ml of xylene, cover the tube tightly, and vortex for 10 seconds.
[0150] b. Centrifuge at 12,000 rpm (~13,400×g) for 2 minutes, carefully aspirate and discard the supernatant, and be careful not to aspirate the sediment.
[0151] c. Add 1ml of absolute ethanol, vortex and mix well. Centrifuge at 12,000 rpm for 2 minutes, discard the supernata...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com