Nested asymmetric PCR reagent kit for detecting alpha 2 globin gene point mutation
A globin and kit technology, which is applied in the field of nested asymmetric PCR melt-linking fluorescence analysis detection kits for α2 globin gene point mutation, can solve the problems of cumbersome operation, time-consuming and labor-intensive, and small detection throughput.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0076] Nested asymmetric PCR kit for detecting point mutations of α2 globin genes WS, QS, CS, CD30, CD31
[0077] 1. Kit composition
[0078] (1) A pair of primers that specifically amplify the full sequence of the α2 globin gene
[0079] F: 5'-gctcgcgggccggcact-3' (SEQ NO. 1),
[0080] R: 5'-ctccattgttggcacattccgggatag-3' (SEQ NO. 2);
[0081] (2) Two DNA single-stranded primers for amplifying the α2 globin gene sequence
[0082] The single-stranded primer sequence for amplifying the target fragment of CD30 / CD31 is: 5'-ctgcggggagaagcagagt-3' (SEQ NO.3),
[0083] The single-stranded primer sequence for amplifying the WS / QS / CS target fragment is: 5'-gggcaggaggaacggctac-3' (SEQ NO.4);
[0084] (3) Probe
[0085] Fluorescent probes that specifically detect CD30 / CD31 wild-type and CD31 (AGG>AAG) mutant types:
[0086] 5'-Cy5-ccctggagaggtgaggctaaa-3' (SEQ NO. 5);
[0087] Quencher probes that specifically detect CD30 / CD31 site wild-type and CD31 (AGG>AAG):
[0088] 5'-aatggcg...
Embodiment 2
[0130] Using the kit and method of Example 1 to detect point mutations of α-thalassemia globin genes WS, QS, CS, CD30, and CD31:
[0131] 1. Sample source and type:
[0132] Select gDNA specimens of confirmed α-thalassaemia from the sample bank of our laboratory, and the genotypes are α CD30 α / αα, α CD30 α / α CD30 α, α CD31 α / αα, α CD31 α / α CD31 1 each of α, α CS α / αα, α CS α / α CS α, α WS α / αα, α WS α / α WS α, α QS α / αα, α QS α / α QS α, -- SEA / α WS α, -- SEA / α QS α, -- SEA / α CS 2 copies of α, αα / αα each, dilute gDNA samples to 100-200ng / μl with sterilized double distilled water for later use.
[0133] 2. Sample detection:
[0134] Apply the above-mentioned reaction system of this kit (Table 1) to the above-mentioned gDNA specimen to be tested, and use the PCR reaction program of the present invention (pre-denaturation at 95°C for 8 minutes; 95°C for 30sec+72°C for 60sec, 30 cycles; 95°C for 30sec+ 63°C 20sec, 40 cycles; 72°C extension for 5min; 95°C denat...
PUM
Property | Measurement | Unit |
---|---|---|
melting point | aaaaa | aaaaa |
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