DNA isolation method and kit
AssigneeEppendorf 5 Prime, Inc.
InventorsE. Kurt Lienau, J. Michael Hurley
Life of this patent
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Abstract
A method and kit for isolating nucleic acids from a nucleic acid containing starting material is disclosed, where the nucleic acids are released from the starting material and precipitated onto a trapping membrane. The method and kit may be used in the context of isolating genomic DNA from blood and isolating BACs from transformed bacterial cultures.
Claims42
A method for isolating nucleic acid from a nucleic acid containing starting material, comprising:
- mixing the starting material with a lysing and denaturing substance for release of the nucleic acid from the starting material;
- separating the mixture into a first portion and a second portion, the first portion having a greater amount of the nucleic acid than the second portion;
- adding an alcohol and detergent substance to the first portion of the mixture to precipitate the nucleic acid onto a nylon trapping membrane;
- washing the nylon trapping membrane to remove nucleic acid and trapping membrane associated contaminants;
- and
- adding a re-suspension buffer to the nylon trapping membrane to rapidly re-suspend and release the nucleic acid from the nylon trapping membrane.
A kit for isolating nucleic acid from a nucleic acid containing starting material comprising:
- a lysing and denaturing substance in an amount sufficient to release the nucleic acid from the starting material;
- an alcohol and detergent substance in an amount sufficient to precipitate the nucleic acid from the starting material;
- and
- a nylon membrane in an amount sufficient to trap the precipitated nucleic acid.
A method for isolating bacterial artificial chromosomes from a bacterial artificial chromosome containing cell culture starting material, comprising:
- mixing the starting material with a lysis substance for release of the bacterial artificial chromosomes from the cell culture;
- passing the mixture through a pre-filter to remove cell debris from the bacterial artificial chromosomes;
- adding an alcohol and detergent substance to the bacterial artificial chromosome containing filtrate to precipitate the bacterial artificial chromosomes onto a trapping membrane;
- washing the trapping membrane with a purification solution to remove bacterial chromosome associated contaminants from the trapping membrane;
- and
- re-suspending the isolated bacterial artificial chromosome in a re-suspension buffer.
Dependencies are inferred from the claim text.