In vitro sorting method
First Claim
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1. A method for increasing the concentration of a nucleic acid molecule by polymerase chain reaction amplification comprising the steps of:
- (a) forming aqueous microcapsules from a thermostable water-in-oil emulsion, wherein a plurality of the microcapsules include a nucleic acid molecule, a bead, and an aqueous solution comprising components necessary for polymerase chain reaction amplification;
(b) amplifying the nucleic acid molecule in the microcapsules to form amplified copies of the nucleic acid molecule which bind to said bead; and
(c) breaking the microcapsules and selecting for said amplified copies of the nucleic acid molecule using a tag, wherein the tag comprises one member of a binding pair.
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Abstract
The invention describes a method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising of the steps of: (a) compartmentalising genetic elements into microcapsules; (b) expressing the genetic elements to produce their respective gene products within the microcapsules; (c) sorting the genetic elements which produce the gene product having a desired activity. The invention enables the in vitro evolution of nucleic acids by repeated mutagenesis and iterative applications of the method of the invention.
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Citations
11 Claims
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1. A method for increasing the concentration of a nucleic acid molecule by polymerase chain reaction amplification comprising the steps of:
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(a) forming aqueous microcapsules from a thermostable water-in-oil emulsion, wherein a plurality of the microcapsules include a nucleic acid molecule, a bead, and an aqueous solution comprising components necessary for polymerase chain reaction amplification; (b) amplifying the nucleic acid molecule in the microcapsules to form amplified copies of the nucleic acid molecule which bind to said bead; and (c) breaking the microcapsules and selecting for said amplified copies of the nucleic acid molecule using a tag, wherein the tag comprises one member of a binding pair. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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Specification