Process for extracting nucleic acid
OwnerHui Cheninventor
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Abstract
The invention provides a process and kit for isolating and purifying nucleic acids such as DNA or RNA or a hybrid molecule of DNA and RNA. A siliceous material combined with a solution is used to prepare highly pure nucleic acids, especially DNA. The siliceous material is a support for absorbing target materials, and the solution according to this invention promotes the target materials to bind to the siliceous material, especially to promote DNA to bind to the siliceous material. The solution is an acid and potassium ion-containing aqueous solution. The invention further provides DNA prepared by using the process. Because no chaotropic agents or other poisonous or costly agents are used in the process, DNA prepared by the process may be used widely, especially in food industry and pharmaceutical industry.
Claims28
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A method for separation and purification of nucleic acids, said method comprising:
- (a) adding an appropriate amount of acidic aqueous solution containing potassium ions into a raw biomaterial containing nucleic acids;
- (b) mixing said acidic aqueous solution containing potassium ions with said raw biomaterial, wherein a concentration of said potassium ions in a mixed binding solution is in a range of 0.3M to saturated, and a pH of the mixed binding solution is in a range of 2.0-4.0;
- wherein said potassium ions are selected from the group consisting of K2SO4, KNO3, KCl, potassium acetate and a mixture thereof;
- and wherein said acidic aqueous solution is selected from the group consisting of acetic acid, propionic acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and a mixture thereof;
- (c) adding a silicon-containing material to said mixed binding solution, wherein said nucleic acids in said raw biomaterial are capable of binding to said silicon-containing material;
- (d) separating nucleic acids bound with said silicon-containing material from impurities;
- and
- (e) separating and eluting nucleic acids from said silicon-containing material.
A kit used for a DNA separation and purification, said kit comprising:
- (a) an acidic aqueous solution containing potassium ions, wherein said acidic aqueous solution is selected from the group consisting of acetic acid, propionic acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and a mixture thereof;
- wherein said potassium ions are selected from the group consisting of K2SO4, KNO3, KCl, potassium acetate and a mixture thereof;
- and wherein a concentration of said potassium ions in a mixed binding solution is adjusted to be in a range of 0.3M to saturated, and a pH of the mixed binding solution is adjusted to be in a range of 2.0-4.0;
- (b) a silicon-containing material that is capable of binding to said DNA;
- and
- (c) an introduction manual listing contents of said kit and providing procedures for said DNA separation or purification.
A method of extracting plasmid DNA from cultured bacteria, said method comprising:
- (a) providing a bacteria lysate by separating and lysing the bacteria from cultured media;
- (b) providing a mixed binding solution by thoroughly mixing an appropriate amount of acidic aqueous solution containing potassium ions with the bacteria lysate, wherein a concentration of said potassium ions in said mixed binding solution is in a range of 0.3M to saturated, and a pH of the mixed binding solution is in a range of 2.0-4.0;
- wherein said potassium ions are selected from the group consisting of K2SO4, KNO3, KCl, potassium acetate and a mixture thereof;
- and wherein said acidic aqueous solution is selected from the group consisting of acetic acid, propionic acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and a mixture thereof;
- (c) adding a silicon-containing material to said mixed binding solution, wherein said plasmid DNA is capable of binding to said silicon-containing material;
- (d) separating said plasmid DNA bound with said silicon-containing material from impurities;
- and
- (e) washing out the impurities and eluting said plasmid DNA from said silicon-containing material to obtain the purified plasmid DNA.
A kit used for extracting plasmid DNA from cultured bacteria, said kit comprising:
- (a) reagents or solutions used for separating and lysing said cultured bacteria;
- (b) an acidic aqueous solution containing potassium ions, wherein said acidic aqueous solution is selected from the group consisting of acetic acid, propionic acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and a mixture thereof;
- wherein said potassium ions are selected from the group consisting of K2SO4, KNO3, KCl, potassium acetate and a mixture thereof;
- and wherein a concentration of said potassium ions in a mixed binding solution of bacterial lysate and said acidic aqueous solution is adjusted to be in a range of 0.3M to saturated, and a pH of said mixed binding solution is adjusted to be in a range of 2.0-4.0;
- (c) a silicon-containing material that is capable of binding to said plasmid DNA;
- (d) reagents or solutions for separating and eluting bound plasmid DNA from impurities and said silicon-containing material;
- and
- (e) an introduction manual listing contents of said kit and providing procedures for extracting said plasmid DNA.
An isolated endogenous nucleic acid free of chaotropic reagent and other toxic reagents, said isolated endogenous nucleic acid is obtainable by a method comprising:
- (a) adding an appropriate amount of acidic aqueous solution containing potassium ions into a raw biomaterial containing said endogenous nucleic acid;
- (b) providing a mixed binding solution by thoroughly mixing said acidic aqueous solution containing potassium ions with said raw biomaterial containing said endogenous nucleic acid, wherein a concentration of said potassium ions in said mixed binding solution is in a range of 0.3M to saturated, and a pH of said mixed binding solution is in a range of 2.0-4.0;
- wherein said potassium ions are selected from the group consisting of K2SO4, KNO3, KCl, potassium acetate and a mixture thereof;
- and wherein said acidic aqueous solution is selected from the group consisting of acetic acid, propionic acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and a mixture thereof;
- (c) adding a silicon-containing material to said mixed binding solution, wherein said endogenous nucleic acid in said raw biomaterial are capable of binding to said silicon-containing material;
- (d) separating silicon-containing material bound endogenous nucleic acid from impurities;
- and
- (e) separating and eluting said endogenous nucleic acid from said silicon-containing material to obtain said isolated endogenous nucleic acid.
An isolated endogenous nucleic acid free of chaotropic reagent and other toxic reagents.
Dependencies are inferred from the claim text.