A virus inactivation, capture and real-time fluorescent constant temperature amplification detection kit and its application
A technology of capturing reagents and kits, which is applied in biochemical equipment and methods, microbe determination/testing, DNA/RNA fragments, etc., and can solve problems such as false positives, low sensitivity, and false negatives in experimental results
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0125] Preparation of inactivated lysate:
[0126] Use deionized water to prepare inactivated lysate according to the following components and concentrations: 0.2M ammonium salt, 2M guanidine isothiocyanate, 0.5M lithium chloride, 20mM ethylenediaminetetraacetic acid, 50mM ethylene glycol bis(2-amino ethyl ether) tetraacetic acid, 0.2% TritonX-100, 0.5% lithium dodecyl sulfate, 50 mM ascorbic acid, 0.5% dithiothreitol, 100 mM HEPES buffer, pH 5.5.
[0127] Add pseudoviruses of known concentration to 200 μL of the above-mentioned inactivated lysate and PBS solution, shake at room temperature for 20 minutes, and use Invitrogen TM Dynabeads supplied TM SILANE Viral NA Kit is verified, and Invitrogen is not added to those who have used the above inactivated lysate TM The lysis reagent in the kit does not use the lysate and lysis steps of the kit, directly extracts and detects with conventional RT-PCR, the results show that there is no significant difference between the two, whic...
Embodiment 2
[0130] Embodiment 2: the synthesis of intermediate probe sequence
[0131] Entrust Sangon Bioengineering (Shanghai) Co., Ltd. to synthesize SEQ ID NO.3 to 11 and the following intermediate probe sequences:
[0132] 5'biotin Oligo(dT) 15 3'
[0133] 5'biotin-C6-Oligo(dT) 25 VN 3'
[0134] 5'NH 2 -C6-Oligo(dT) 30 3'
Embodiment 3
[0135] Example 3: Preparation of capture magnetic beads Capture MB-1
[0136] 10 mg Dynabeads MyOne Streptavidin T1 (from Invitrogen) was washed twice with PBS, resuspended in 500 μL 5X SSC buffer, and 10 μmol 5’biotin Oligo(dT) was added 15 The 3' intermediate probe was reacted at room temperature for 30 minutes, then magnetically separated, and the remaining probe content was detected in the supernatant. The magnetic beads-intermediate probe was washed 5 times with 2X SSCbuffer, and finally stored in PBS buffer containing 0.1% preservatives and 0.05% Tween20. The content of the probe on the above-mentioned magnetic beads was 935 pmol / mg. Add 800 pmol of SEQ ID NO.3 and 800 pmol of SEQ ID NO.4 to the above-mentioned 10 mg magnetic bead-intermediate probe buffer, react at room temperature for 30 minutes, magnetically separate, and detect that the remaining probe content in the supernatant is zero, that is All capture probes have been bound to the surface of the above-mention...
PUM
Property | Measurement | Unit |
---|---|---|
particle diameter | aaaaa | aaaaa |
particle diameter | 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