Method for separation and purification of malignant tumor cell dna by cationic montmorillonite
A cationic, malignant tumor technology, applied in the field of separation and purification of malignant tumor cell DNA, can solve the problems of inconvenient high-throughput operation, single purification model, and high cost of material synthesis, so as to reduce the preparation cost, improve the dispersion effect, and enhance the adsorption. effect of action
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0027] Cell Lysis:
[0028] Culture HeLa cells in a 6cm-diameter petri dish, then add 500 μL of 2% sodium dodecyl sulfate solution to lyse, transfer the cell lysate into an EP tube and heat at 80°C for 5 min, at an amplitude of 20%, Sonicate at a power of 500W and store in a -20°C refrigerator for later use.
[0029] Preparation of lithium-ion montmorillonite:
[0030] Dissolve Li in 9 mL of deionized water 2 CO 3 0.23 g, add 1.00 g of calcium-based montmorillonite, react in a constant temperature water bath at 60 °C for 24 h, centrifuge, dry at 60 °C for 24 h, and store in a dry dish in an airtight manner, marked as 0.5Li-MMT.
[0031] DNA adsorption:
[0032] Take 70 μL of 30 g / L lithium-ion montmorillonite suspension, add 20 μL of cell lysate and 10 μL of sodium acetate solution with a pH of 5.0, react for 2 minutes and centrifuge at a centripetal acceleration of 5000 g for 2 minutes, take the supernatant and wash with agar The DNA content was determined by gel electro...
Embodiment 2
[0036] Cell Lysis:
[0037] Culture HeLa cells in a petri dish with a diameter of 6 cm, then add 500 μL of 2% sodium dodecyl sulfate solution to lyse, transfer the cell lysate into an EP tube and heat at 90°C for 5 min, at an amplitude of 20%, Sonicate at a power of 500W and store in a -20°C refrigerator for later use.
[0038] Preparation of lithium-ion montmorillonite:
[0039] Dissolve Li in 9 mL of deionized water 2 CO 3 0.47 g, add 1.00 g of calcium-based montmorillonite, react in a constant temperature water bath at 60 °C for 24 h, centrifuge, dry at 60 °C for 24 h, and store in a dry dish in an airtight manner, marked as Li-MMT.
[0040] DNA adsorption:
[0041] Take 70 μL of 30 g / L lithium-ion montmorillonite suspension, add 20 μL of cell lysate and 10 μL of sodium acetate solution with a pH of 5.0, react for 2 minutes and centrifuge at a centripetal acceleration of 5000 g for 2 minutes, take the supernatant and wash with agar The DNA content was determined by gel...
Embodiment 3
[0045] Cell Lysis:
[0046] Culture HeLa cells in a 6cm-diameter petri dish, then add 500 μL of 2% sodium dodecyl sulfate solution for lysis, transfer the cell lysate into an EP tube and heat at 100°C for 5 minutes, at an amplitude of 20%, Sonicate at a power of 500W and store in a -20°C refrigerator for later use.
[0047] Preparation of lithium-ion montmorillonite:
[0048] Dissolve Li in 9 mL of deionized water 2 CO 30.71 g, add 1.00 g of calcium-based montmorillonite, react in a constant temperature water bath at 60 °C for 24 h, centrifuge, dry at 60 °C for 24 h, and store in a dry dish in an airtight manner, marked as 1.5Li-MMT.
[0049] DNA adsorption:
[0050] Take 70 μL of 30 g / L lithium-ion montmorillonite suspension, add 20 μL of cell lysate and 10 μL of sodium acetate solution with a pH of 5.0, react for 2 minutes, and then centrifuge at a centripetal acceleration of 5000 g for 2 minutes. The supernatant was taken and the DNA content was determined by agarose g...
PUM
Login to view more Abstract
Description
Claims
Application Information
Login to view more - 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.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap