Nucleic acid purification method

摘要

The present invention relates to a method for purifying a defined amount of nucleic acids from a nucleic acid-containing sample, which has at least the following steps: (a.) contacting the nucleic acid-containing sample with a defined amount of a nucleic acid binding phase with the following features: (i) the nucleic acid binding phase has nucleic acid binding ligands that have at least one protonatable group; (ii) the nucleic acid binding ligands are bound to a carrier; (iii) the nucleic acid binding phase has a surface with a low charge density, wherein the amount of nucleic acids in the sample exceeds the binding capacity of the amount of nucleic acid binding phase used; (b.) binding of the nucleic acids to the nucleic acid binding phase at a pH (binding pH) that is below the pKs value of at least one of the protonatable groups; (c.) elution of the nucleic acids at a pH that is above the binding pH, wherein a defined amount of nucleic acids is obtained. Furthermore, corresponding kits and nucleic acid binding phases, which can be used for the purification of nucleic acids, are disclosed.

主权项

1. A method for isolating and/or purifying a defined amount of nucleic acids from a nucleic acid-containing sample, the method having at least the following steps:
a. contacting the nucleic acid-containing sample with a defined amount of a nucleic acid binding phase with the following features:
(i) the nucleic acid binding phase has nucleic acid binding ligands, which have at least one protonatable group;(ii) the nucleic acid binding ligands are bound to a carrier, wherein the low charge density is achieved by at least one of the following features:
(aa) the nucleic acid binding ligands have, bound to the carrier, in each case not more than one or two protonatable groups for binding of the nucleic acids, and/or(bb) the nucleic acid binding ligands are selected from the group of mono- and diamines, and/or(cc) the carrier is coated with a mixture of nucleic acid binding ligands and diluting groups, wherein the proportion of nucleic acid binding ligands relative to the diluting groups is ≦50%; and/or(dd) the nucleic acid binding ligands bound to the carrier are in deficit, such that ≦50% of functional groups or groups functionalizable with nucleic acid binding ligands on the carrier are functionalized with nucleic acid binding ligands;(iii) the nucleic acid binding phase has a surface with a low charge density, wherein the amount of nucleic acids in the sample exceeds the binding capacity of the amount of nucleic acid binding phase used; b. binding of the nucleic acids to the nucleic acid binding phase at a pH which is a binding pH that is below the pKs value of at least one of the protonatable groups; c. elution of the nucleic acids at a pH that is above the binding pH, wherein a defined amount of nucleic acids is obtained.