Method for purifying a target nucleic acid

摘要

The present invention pertains to method for purifying at least a target nucleic acid from a sample, said method comprising at least the following steps: a) incubating the sample with at least one protein-degrading compound; b) binding the target nucleic acid to a solid phase; c) eluting the target nucleic acid from the solid phase; d) incubating the eluted target nucleic acid with at least one protein-degrading compound; e) binding the target nucleic acid again to a solid phase; f) optionally eluting the bound target nucleic acid from the solid phase. It was surprisingly found that performing a second protein digestion step after the target nucleic acid was bound and eluted from a solid phase before the nucleic acids are rebound to a solid phase is very efficient in reducing remaining protein contaminations in the isolated nucleic acid.

主权项

1. A method for purifying at least one target nucleic acid from a sample, comprising:
(A) incubating the sample with at least one protein-degrading compound, (B) binding the target nucleic acid to a first solid phase, (C) eluting the target nucleic acid from the first solid phase, (D) incubating the eluted target nucleic acid with at least one protein-degrading compound, and (E) binding the target nucleic acid again to a second solid phase, wherein the protein degrading compound of steps (A) and (D) is a proteolytic enzyme; and wherein binding in step (B), step (E), or both steps (B) and (E) are performed in the presence of an alcohol at a concentration of at least 30% v/v and at least one chaotropic agent.