Method for rapid purifiction of nucleic acids using layered ion-exchange membranes
AssigneeMillipore Corporation
InventorsYolanda Nieuwkerk, Robert J. Barry, Malcolm G. Pluskal, Richard A. Hamilton
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Abstract
The present invention relates to a device method and kit for the convenient and rapid isolation and purification of nucleic acids, proteins, peptides, carbohydrates and oligosaccharides from heterogeneous biological samples. The device comprises a membrane assembly comprised of layers of microporous, polymeric membranes functionalized with ion-exchange groups. The device is reusable for like samples, relatively inexpensive compared to currently available separation techniques and is disposable, thereby avoiding cross-contamination of biological samples.
Claims9
A method for the rapid isolation and purification of a nucleic acid of interest from a cell lysate sample containing the nucleic acid of interest using ion-exchange membranes, comprising the steps of:
- a) applying a buffered solution of sample containing nucleic acid to a membrane assembly which contains a plurality of stacked microporous membranes with pore sizes of 0.1 to 12 microns functionalized with ion-exchange groups capable of adsorbing ionized nucleic acids thereto, wherein the nucleic acid is retained by the membrane;
- b) washing the membrane assembly with an elution buffer of a determined ionic strength sufficient to release weakly adsorbed elute contaminating substances contained in said sample from the membrane;
- c) washing the membrane assembly with an elution buffer of determined ionic strength sufficient to release the bound nucleic acid from the membrane;
- and
- d) recovering the purified nucleic acid.
Dependencies are inferred from the claim text.