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US 10,883,100 B2Granted
Devices and methods for plasmid purification
AssigneePhyNexus, Inc.
InventorsChris Suh, Carrie Loan Kim Huynh, Lee Hoang, Douglas T. Gjerde, Jonathan Michael Grambow
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Abstract
The invention provides columns (including pipette tip columns) and automated methods for the purification of nucleic acids including plasmids. Nucleic acids can be purified from unclarified, clarified or partially-clarified cell lysates that contain cell debris. The columns typically include a bed of medium positioned above a bottom frit and with an optional top frit. Plasmid preparation scales include miniprep, midiprep, maxiprep, megaprep and gigaprep.
Claims8
An automated method for large scale plasmid preparation, the method comprising:
- (a) providing a cell lysate including particulates, liquid, and plasmid DNA, the particulates including cell debris;
- (b) providing a column, wherein the column is capable of plasmid DNA capture;
- (c) providing a filter;
- (d) passing the cell lysate through the filter to partially remove the particulates from the cell lysate to produce a filtrate in a single filtration step, such that
- a first portion of the particulates is removed from the cell lysate by the filter, and
- the filtrate includes the plasmid DNA and a second portion of the particulates that passes freely through the filter;
- (e) passing the filtrate through the column, such that a portion of the plasmid DNA in the filtrate is captured on the column;
- (f) passing a wash solution through the column;
- and
- (g) eluting the plasmid DNA by passing a desorption solution through the column,
- wherein an amount of plasmid DNA eluted from the column is at least 750 pg,
- wherein the passing the filtrate through the column, the passing the wash solution through the column, and the eluting the plasmid DNA are performed at predetermined times.
9 more elements
Dependencies are inferred from the claim text.