News Column

Patent Application Titled "Nucleic Acid Isolation and Purification System" Published Online

February 6, 2014



By a News Reporter-Staff News Editor at Politics & Government Week -- According to news reporting originating from Washington, D.C., by VerticalNews journalists, a patent application by the inventor PERRY, MICHAEL P. (Downingtown, PA), filed on July 12, 2012, was made available online on January 23, 2014.

The assignee for this patent application is E I Du Pont De Nemours And Company.

Reporters obtained the following quote from the background information supplied by the inventors: "Isolation and purification of nucleic acids is important for clinical diagnostics and basic molecular and microbiology research such as public health genetic testing, forensics, phylogenetics, transgenics, gene sequencing, and metabolic pathway analysis.

"Nucleic acid purification is typically performed via anion exchange on positively charged substrates, or chaotropic capture on silica substrates. A commonly used nucleic acid binding material is a glass surface due to its ability to reversibly bind nucleic acids in the presence of chaotropic reagents and/or alcoholic additives (Boom et al. (1990) J. Clin. Microbiol. 28:495-503). In this type of binding and elution, DNA is 'bound' to the negatively charged substrate from very high ionic strength solution (e.g. guanidine thiocyanate), and subsequently eluted in a significantly lower ionic strength solution. These substrates are typically silica fiber filter discs or ion exchange resins that are contained in column type devices that use gravity flow, centrifugation, suction, or pressure from a plunger for nucleic acid isolation. In these types of devices, particulates and/or viscous material (such as extracellular polysaccharides) in a complex sample tend to clog the filter or resin.

"WO2011017251 discloses a process for collection of nucleic acids of microorganisms that involves a fibrous nucleic acid binding surface in a chamber where expansion may occur, and compression of the fibrous nucleic acid binding surface to expel fluids.

"Many nucleic acid isolation devices and kits that are available require lengthy procedures that include multiple steps and reagents, long incubation times, and complex devices. For example, WO2010075116 discloses a clarification/binding device comprising a clarification column and a binding column, and a vacuum manifold.

"There is thus a need for a simple nucleic acid isolation device that can handle relatively large volumes, takes a minimal amount of time, does not clog with particulate or viscous samples, and can be used in a rapid nucleic acid isolation process."

In addition to obtaining background information on this patent application, VerticalNews editors also obtained the inventor's summary information for this patent application: "The invention relates to a device can be used for rapid nucleic acid isolation from relatively large volume samples with minimal clogging.

"Accordingly, the invention provides a multi-component device for isolating nucleic acids from a sample comprising: a) a cylindrical upper chamber and a cylindrical lower chamber, said upper and lower chambers detachably connectable at first ends, with said connection affording in its interior a smooth, uniform transition between said upper and lower chambers and at its exterior on the lower chamber a flange, and said lower chamber comprising a screen at its second end; b) a fibrous nucleic acid binding surface partially filling the inside of the upper and lower chambers and maintained in the chambers by the screen; c) a removable cap at the second end of the upper chamber and a removable cap at the second end of the lower chamber; d) a first collection tube into which the lower chamber slides up to the flange wherein an air-tight seal is produced; and e) an optional second collection tube into which the lower chamber partially slides.

"In another embodiment the invention provides a nucleic acid purification kit comprising a device comprising: a) a cylindrical upper chamber and a cylindrical lower chamber, said upper and lower chambers detachably connectable at first ends, with said connection affording in its interior a smooth, uniform transition between said upper and lower chambers and at its exterior on the lower chamber a flange, and said lower chamber comprising a screen at its second end; b) a fibrous nucleic acid binding surface partially filling the inside of the upper and lower chambers and maintained in the chamber by the screen; c) a removable cap at the second end of the upper chamber and a removable cap at the second end of the lower chamber; d) a first collection tube into which the lower chamber slides up to the flange wherein an airtight seal is produced; and e) optionally a second collection tube into which the lower chamber partially slides.

"In yet another embodiment the invention provides a method of isolating nucleic acids comprising: a) forming a binding mixture comprising: i) a sample containing nucleic acids; and ii) a nucleic acid binding solution; wherein the binding mixture is either formed within or is added to a double-chamber device comprising: iii) a cylindrical upper chamber and a cylindrical lower chamber, said upper and lower chambers detachably connectable at first ends, with said connection affording in its interior a smooth, uniform transition between said upper and lower chambers and at its exterior on the lower chamber a flange, and said lower chamber comprising a screen at its second end; iv) a fibrous nucleic acid binding surface partially filling the inside of the upper and lower chambers and maintained in the chambers by the screen; and v) a removable cap on the second end of the lower chamber; b) incubating the binding mixture in the double-chamber device to allow nucleic acid binding; c) removing the cap and draining the unbound sample by gravity flow; d) placing the double-chamber device in a first collection tube wherein an air-tight seal is produced; e) applying a wash solution to the double-chamber device; f) releasing the seal to drain the wash solution; g) separating the chambers of the double-chamber device; h) placing the lower chamber in a centrifuge tube; i) adding elution buffer to release bound nucleic acid; and j) centrifuging to collect the elution buffer containing nucleic acid from the sample; wherein no filtering, plunging, or vacuum step is used.

BRIEF DESCRIPTION OF THE FIGURES

"FIG. 1 shows diagrams of a two-chamber fiber-containing device showing separate components in (A) and a set of assembled components in (B).

"FIG. 2 shows diagrams of types of connections to join upper and lower chambers of the two-chamber fiber-containing device: with threads on the upper and lower chambers that screw into a collar as separate components (A) and assembled (B); or with threads on the upper chamber that screw into the lower chamber as separate components (C) and assembled (D).

"FIG. 3 shows diagrams of a prototype two-chamber fiber-containing device that uses a modified 3-mL syringe and a modified spin column with original syringe and spin column in (A), modified syringe and spin column in (B), and a set of assembled components, with additional collection microcentrifuge tube in (C)."

For more information, see this patent application: PERRY, MICHAEL P. Nucleic Acid Isolation and Purification System. Filed July 12, 2012 and posted January 23, 2014. Patent URL: http://appft.uspto.gov/netacgi/nph-Parser?Sect1=PTO2&Sect2=HITOFF&u=%2Fnetahtml%2FPTO%2Fsearch-adv.html&r=1618&p=33&f=G&l=50&d=PG01&S1=20140116.PD.&OS=PD/20140116&RS=PD/20140116

Keywords for this news article include: E I Du Pont De Nemours And Company.

Our reports deliver fact-based news of research and discoveries from around the world. Copyright 2014, NewsRx LLC


For more stories covering the world of technology, please see HispanicBusiness' Tech Channel



Source: Politics & Government Week


Story Tools