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Free DNA extraction and enrichment kit with guanidine hydrochloride 2019-07-05 13:57:25


Nucleic acids include deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), and are a class of biological macromolecules with genetic information. The study found that free DNA also contains very important genetic information. Therefore, in recent years, free DNA research has become a hot field in genomic molecular diagnostic research, and has been gradually applied to all stages from the initial diagnosis of tumor to treatment and development. However, the extraction technique of free DNA still needs further improvement.


The researchers developed a free DNA extraction and enrichment kit and a method for extracting free DNA, aiming to solve the technical problem that the organic solvent has certain toxicity in the existing free nucleic acid extraction method.


Free DNA extraction and enrichment kit

The kit includes sample lysate, binding solution, primary wash, secondary wash, eluent, and nanomagnetic beads:


  • The sample lysate components include nuclease-free water, reagent A and reagent B. Reagent A is at least one of guanidinium isothiocyanate, guanidine hydrochloride, sodium iodide and sodium perchlorate. Reagent B is at least one of Tween20, Triton-X100, SDS, NP40 and a polyoxyethylene type nonionic surfactant;
  •  In addition to the components of the sample lysate, binding solution also include reagent C, which is at least one of isopropanol and absolute ethanol;
  • The composition of the primary wash solution is the same as the combined solution;
  •  The second washing liquid is one of nuclease-free water, a molar concentration of 0.1~1 M sodium chloride solution, Tris buffer or TE buffer;
  • The eluent is one of nuclease-free water, Tris buffer or TE buffer;
  • The magnetic core of nano-magnetic beads is Fe-Co-NiOX. The particle size distribution is between 300-400nm. The surface layer of the magnetic core is coated with at least carbon layer and inorganic silica layer from inside to outside. And polyethylene glycol is covalently grafted on the outermost layer.



Free DNA extraction method


  • Add the protease K first, then add the sample, and the lysate sequence, add the components in the centrifuge tube, shake and incubate to carry out sample lysis;
  • The nano magnetic beads and the binding solution are proportioned to form a nano magnetic bead binding solution;
  • Add the prepared nano magnetic bead binding solution to the sample, shake and mix, and discard the supernatant;
  • Wash the free DNA with a wash solution;
  • Wash the free DNA with a secondary washing solution;
  • Add the eluent and shake to resuspend the nanobeads and further shake to elute the free DNA on the nanobeads.


The method do not use toxic reagents such as benzene and chloroform in the traditional method to minimize the damage to the experimental operators, fully comply with the modern environmental protection concept, and better meet the clinical use conditions; Lysis, denaturation and promotion of nucleic acid enrichment, most samples do not require additional use of proteinase K and sample lysate; It takes less time and is easy to operate for all binding, washing and elution processes at room temperature.


Reference

Free DNA extraction and enrichment kit and free DNA extraction method. CN109913447A, 2019-06-21.


Edited by Suzhou Yacoo Science Co., Ltd.

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