Application of Tetraphenylethylene Derivatives in Detection of Methyltransferase Activity and Methyltransferase Inhibitor Concentration
A technology of methyltransferase and tetraphenylethylene, which is applied in the biological field to achieve the effects of high sensitivity, good water solubility and simple synthesis
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0054] A short-chain nucleic acid modified with the quencher molecule dimethylaminoazobenzoyl at both ends, as shown in SEQ ID No: 1 (5'-GTTGGGATCGAGAG-3'), and a long-chain nucleic acid, such as SEQ ID No: 4 ( 5'-AGTGACATGATTTCCTCTCGATCCCAACCGCCGTATAGATAG-3'), mixed in an aqueous solution to obtain a quenched molecularly modified double-stranded nucleic acid (dsDNA3-2Q);
[0055] In a total reaction system of 50 μL, 2 μM dsDNA3-2Q, 1 μL enzyme reaction buffer solution [10×buffer: 200 mMTris-HAc, 500 mMKAc, 100 mMMg(Ac)2, 10 mMDTT, pH7.9], 80 μS-adenosylmethionine Acid (SAM), 400U / mL restriction endonuclease (DpnI) and different concentrations of methyltransferase Dam (concentrations were 0, 0.25, 0.5, 1, 2.5, 5, 10, 20, 40, 80 and 160U / mL), placed at 37°C for 5 hours, and then placed in a 90°C water bath for 10 minutes to inactivate Dam and DpnI, and finally cooled to room temperature to obtain a mixed solution;
[0056] The above mixed solution was mixed evenly with 50 μL ...
Embodiment 2
[0061] A short-chain nucleic acid modified with a quencher molecule dimethylaminoazobenzoyl at both ends, as shown in SEQ ID No: 2 (5'-GTTGGCCGGGAGAG-3'), and a long-chain nucleic acid, such as SEQ ID No: 5 ( 5'-AGTGACATGATTTCCTCTCCCGGCCAACCGCCGTATAGATAG-3'), mixed in an aqueous solution to obtain a double-stranded nucleic acid (dsDNA4-2Q) modified by quenching molecules;
[0062] In a total reaction system of 50 μL, mix 2 μM dsDNA4-2Q, 1 μL enzyme reaction buffer solution [10×buffer: 200 mMTris-HAc, 500 mMKAc, 100 mMMg(Ac)2, 10 mMDTT, pH7.9], 80 μS-adenosylmethionine Acid (SAM), 400U / mL restriction endonuclease (HpaII) and different concentrations of methyltransferase HpaIIMTase (concentrations were 0, 0.25, 0.5, 1, 2.5, 5, 10, 20, 40, 80 and 160U / mL), placed at 37°C for 5 hours, and then placed in a 90°C water bath for 10 minutes to inactivate HpaIIMTase and HpaII, and finally cooled to room temperature to obtain a mixed solution;
[0063] The above mixed solution was unif...
Embodiment 3
[0068] A short-chain nucleic acid with a quencher molecule dimethylaminoazobenzoyl modified at both ends, as shown in SEQ ID No: 3 (5'-GTTGGCGCGGAGAG-3'), and a long-chain nucleic acid such as SEQ ID No: 6 (5 '-AGTGACATGATTTCCTCTCCGCGCCAACCGCCGTATAGATAG-3'), mixed in an aqueous solution to obtain a double-stranded nucleic acid (dsDNA5-2Q) modified by quenching molecules;
[0069] In a total reaction system of 50 μL, 2 μM dsDNA5-2Q, 1 μL enzyme reaction buffer solution [10×buffer: 200 mMTris-HAc, 500 mMKAc, 100 mMMg(Ac)2, 10 mMDTT, pH7.9], 80 μS-adenosylmethionine acid (SAM), 400U / mL restriction endonuclease (BstUI) and different concentrations of methyltransferase M.SssI (concentrations were 0, 0.25, 0.5, 1, 2.5, 5, 10, 20, 40, and 160U / mL), placed at 37°C for 5 hours, and then placed in a 90°C water bath for 10 minutes to inactivate M.SssI and BstUI, and finally cooled to room temperature to obtain a mixed solution;
[0070]Mix the above mixed solution with 50 μL of an aqueo...
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