One-step Wash Magnetic Bead Method Saliva DNA Extraction Kit

Abstract

The invention belongs to the technical field of molecular biology detection. Specifically, the application provides a magnetic bead method saliva DNA extraction kit that only needs one-step washing, which includes a lysis solution, a binding solution, a proteinase K solution, a washing solution and an elution solution. buffer. A saliva DNA extraction kit includes lysing solution, binding solution, proteinase K solution, washing solution and elution buffer. The lysate contains Tris, EDTA, NaCl, NaDC, dodecyl-N-betaine, NLS, GuHCl, TritonX100, NH 4 Cl, mercaptoethanol; binding solution including isopropanol, PEG 8000, Brij 58; washing solution including GuHCl, Tris, ethanol, sodium bicarbonate; elution buffer including Tris, EDTA. In the present application, by preparing a washing solution containing a relatively high concentration of sodium bicarbonate (slightly stronger than sodium acetate and sodium chloride), the ion balance and the effect of providing alkalinity are taken into account at the same time, and a zwitterionic detergent is used in combination. Alkyl-N-betaine, a magnetic bead-based saliva DNA extraction kit was constructed that only required one-step washing.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection. Specifically, the application provides a magnetic bead method saliva DNA extraction kit that only needs one-step washing, which includes a lysis solution, a binding solution, a proteinase K solution, a washing solution and an elution solution. buffer. Background technique [0002] Nucleic acid extraction is the basis of various molecular biology detection methods, and efficient and complete DNA extraction is the basis for PCR amplification, library construction, and sequencing. In addition to tissue and organ samples, the most commonly used in vitro samples in molecular biology testing are blood samples, but blood collection requires professionals to use sterile blood collection equipment, which is costly and complicated. Immediate pain and fear are rejected by many people; for these reasons, molecular biology testing with blood samples is inconvenient in many situations, esp...

AI Technical Summary

Problems solved by technology

Currently known phenol chloroform method, salting-out method, adsorption column method, immunoaffinity method, magnetic bead method and other existing DNA extraction methods that can be used for saliva DNA extraction mainly include adsorption column method and magnetic bead method. There are obvious deficiencies in the extraction effect and the concentration of the extracted DNA, and the magnetic bead method is easy to operate (such as the magnetic bead method of CN201611240409, CN201810169271, CN201610848178, CN201110105238, etc.) in the extraction of DNA by the magnetic bead method. , not only consumes time, but also requires more delicate operation when absorbing the supernatant after washing, which is prone to pollution / not suitable for on-site operation outside the laboratory) and the extraction effect also has a lot to be improved. It is necessary to develop a method that is easier to operate Advanced magnetic bead DNA extraction method to improve extraction efficiency / meet the needs of on-site rapid sample processing in screening, forensic investigation, etc.

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