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DNA (desoxyribonucleic acid) eluent and elution method

A deoxyribonucleic acid and eluent technology, which is applied in the field of separation and purification of deoxyribonucleic acid, can solve the problems of inability to effectively overcome gravity and low elution capacity

Inactive Publication Date: 2014-05-28
BEIJING NORMAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned method of desorbing DNA with a high-salt solution cannot effectively overcome the attraction of positively charged groups on the surface of the DNA and the solid adsorbent, and the elution capacity is low.

Method used

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  • DNA (desoxyribonucleic acid) eluent and elution method
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  • DNA (desoxyribonucleic acid) eluent and elution method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Deoxyribonucleic acid (DNA) adsorption: 3 microliters of DL2000 DNA Marker with a concentration of 90μg / mL and 5 microliters of 4g / L dendrimer-modified silica nanomaterial (SNP-PAMAM) as solid-phase adsorption materials Add a 200 μl centrifuge tube, shake for 10 min at room temperature, stand for 10 min at 4° C., centrifuge at 3500 rpm for 10 min, and collect the SNP-PAMAM-DNA complex.

[0035] The residual DNA content in the supernatant was detected with a fluorescence spectrophotometer (Cary Eclipse, Varian, Palo Alto, CA, USA) using ethidium bromide as a fluorescent probe. The specific method is as follows:

[0036] Add 1.5 microliters of 1mg / mL ethidium bromide (EB) solution to a 5mL graduated test tube, add three times distilled water to make up to the mark, shake up, and excite at 510nm wavelength on a fluorescence spectrophotometer and emit at 620nm wavelength ( The emission and excitation slits are both 10nm, the thickness of the fluorescent cell is 1cm), and the fl...

Embodiment 2

[0043] Add 4 microliters of herring sperm DNA (100μg / mL) and 5 microliters of 4g / L dendrimer modified silica nanomaterials (SNP-PAMAM) as solid-phase adsorption materials into a 200 microliter centrifuge tube, and shake at room temperature After 10 minutes, centrifuge for 10 minutes at 3500 rpm to collect the SNP-PAMAM-DNA complex.

[0044] The residual DNA content in the supernatant was detected with a fluorescence spectrophotometer (Cary Eclipse, Varian, Palo Alto, CA, USA) using ethidium bromide as a fluorescent probe, and the amount of DNA bound to the solid-phase adsorbent was calculated.

[0045] Add the collected SNP-PAMAM-DNA complex to a centrifuge tube containing 15 microliters of eluent. The eluent contains buffer components of 1mM phosphoric acid, 3mM sodium hydroxide and 2mM sodium tetraborate and 5mM [ C 8 MIM]BF 4 Ionic liquid, and the pH of the eluent is 10.8. After shaking for some time at room temperature, centrifuge at 3500 rpm for 10 min, collect the supernatan...

Embodiment 3

[0048] The DNA used in this embodiment is DL2000 DNA Marker (with a concentration of 90 μg / mL). Unless otherwise specified, the operating parameters of this embodiment are the same as those of Embodiment 1.

[0049] The DNA adsorption process uses 3 microliters of DL2000 DNA Marker (with a concentration of 90 μg / mL), and undergoes adsorption, elution, and separation and detection by capillary electrophoresis-UV technology.

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Abstract

The invention provides a DNA (desoxyribonucleic acid) eluent. The eluent comprises an acid-base buffer solution and an ionic liquid, wherein the ionic liquid is a compound containing imidazolyl cations; the pH value of the eluent is 5 to 13. The eluent provided by the invention comprises the compound containing imidazolyl cations, wherein the compound interacts with negatively charged DNA; the DNA is eluted off from the surface of a solid phase adsorbing material; based on the pH value of the eluent, the electrostatic attraction between the solid phase adsorbing material and DNA fragments is lowered and the elution capability of the eluent is further improved. The invention further provides a DNA elution method adopting the eluent for adsorbing the solid phase adsorbing material.

Description

Technical field [0001] The present invention relates to the technical field of separation and purification of deoxyribonucleic acid, in particular to an eluate and an elution method for eluting deoxyribonucleic acid. Background technique [0002] Solid phase extraction is a physical extraction process that includes liquid phase and solid phase. During the extraction process, the adsorption force of the solid relative to the analyte is greater than the sample base liquid. When the sample contacts the solid phase adsorbent, the analyte is adsorbed on the solid phase Adsorb the surface of the material, and then use an appropriate solvent to elute the analyte. The solid-phase adsorption material in this application is insoluble and has a matrix that can interact with the phosphate groups of the nucleic acid backbone. The solid-phase adsorption material can be in the form of porous or non-porous particles, powder particles or fibers, and can be nano-sized or micro-sized, and the surf...

Claims

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Application Information

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IPC IPC(8): C07H1/06C07H21/04
Inventor 秦卫东努尔古丽·拉提莆
Owner BEIJING NORMAL UNIVERSITY
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