Use of functionalised porous membranes or matrices for purification of nucleic acids and corresponding methods
AssigneeEppendorf AG
InventorsThomas Kolzau, Heinz Gerhard Köhn, Wilhelm Plüster, Mathias Ulbricht
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Abstract
Application of a functionalized porous membrane to purify nucleic acids, the binding properties of said membrane with respect to nucleic acids being adjustable by controlling the conditions of an ambient medium, the membrane being functionalized by deprotonated groups, and corresponding methods for purifying nucleic acids.
Claims26
Using a functionalized porous membrane to purify nucleic acids, said membrane's binding properties with respect to nucleic acids being adjustable by controlling the conditions in an ambient medium,
- characterized in that
- the membrane is functionalized with deprotonatable groups.
Application as claimed in one of the above claims, characterized in that the membrane is made of plastic.
Application as claimed in one of the above claims, characterized in that the diameter of the membrane's pores is between 0.2 and 10 μ.
Application as claimed in one of the above claims, characterized in that the membrane's thickness is less than 500 μ.
Application as claimed in one of the above claims, characterized in that the membrane is designed to be used in microtitration filter trays, spin columns or reaction containers.
Purification of nucleic acids by using a flow-permeable matrix which is fitted with a functionalized surface of which the binding properties relative to nucleic acid are adjustable by means of the conditions of an ambient medium,
- characterized in that
- said matrix is functionalized with deprotonatable groups.
Application as claimed in either of claims 11 and 12, characterized in that the matrix is in the form of a non-woven, felt, fibers, sintered material, correspondingly designed pipet tips or reaction containers (consumables) with microstructure ducts.
Application as claimed in one of the above claims, characterized in that the matrix is designed for use in micro-titration filter trays, spin columns or reaction containers.
Method as claimed in one of the above claims, characterized in that an elution buffer, of which the pH value is adjusted to exceed the pK value of the membrane/matrix surface, is allowed to permeate the membrane/matrix in order to elute the bound nucleic acids.
Method as claimed in one of the above claims, characterized in that an elution buffer, of which the ionic concentration has been adjusted to less than 500 mM, is allowed to permeate the membrane/matrix for the purpose of eluting the bound nucleic acids.
Method as claimed in one of the above claims, characterized in that the binding-, washing- and/or elution-buffer is vacuum aspirated through the membrane/matrix.
Method as claimed in one of the above claims, characterized in that the elution of the nucleic acids bound at the membrane/matrix is carried out at room temperature.
Method as claimed in one of the above claims, characterized in that the raw material is subjected to pre-purification in order to remove coarse cell debris and the like.
Method as claimed in the preceding claim, characterized in that the raw material is a clarified cell lysate.
Dependencies are inferred from the claim text.