| 摘要 |
PURPOSE: An improved filtration system is provided and a method for isolating or separating nucleic acids, particularly DNA, from cellular debris is also provided. The system and method are designed to be used in conjunction with both a conventional micro centrifuge and any one of a number of different conventional vacuum manifolds CONSTITUTION: A method for isolating a nucleic acid material using a filter assembly for filtering materials by means of centrifugal force or vacuum assist comprising: a filter basket comprising a cylindrical tube, the filter basket having a first end and a second end, the first end of the fitter basket being open and the second end of the filter basket being adapted to receive and hold a filtration means, the adaptation comprising a perforate structure at the filter basket second end, the perforate structure being sufficiently perforate to support and retain said filter means while permitting fluid to flow there through; and filter means fitted into the filter basket, the filter means being adapted to reversibly bind the nucleic acid material; a vacuum manifold adapter comprising a cylindrical vacuum adapter having a first end and a second end, the first end of the adapter being interiorly configured frictionally to receive the second end of the filter basket, and the second end of the adapter being interiorly configured to couple to a vacuum manifold; and a collection tube having an open end and a closed end, the end of the filter basket adjacent to the filter means fitting into the open end of the collection tube; the method comprising the steps of: a. providing cells containing the nucleic acid material; b. lysing or disrupting the cells, thereby forming a lysate; c. transferring the lysate solution to the filter basket, in the presence of a chaotropic agent capable of promoting binding of the nucleic acid material to the silica; d. removing the lysate solution from the filter basket using removal steps comprising either (i) placing the filter basket in the collection tube and subjecting it to centrifugal force, or (ii) fitting the filter basket onto the first end of the manifold adapter and the second end onto a vacuum manifold and subjecting it to a vacuum; e. washing the filter basket with a wash solution, using centrifugation or a vacuum; and f. eluting the nucleic acid material from the filter basket by adding an elution buffer or water to the basket, placing the basket in a second collection tube and subjecting it to centrifugal force.
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