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US 7,678,554 B2Granted
Nucleic acid shuffling
AssigneePresident and Fellows of Harvard College
InventorsDavid R. Liu, Joshua A. Bittker
Life of this patent
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Abstract
Disclosed is a method of altering a nucleic acid. The method includes fragmenting a parent nucleic acid strand to generate nucleic acid fragments. At least a subset of the fragments are ligated to generate shuffled nucleic acid strands. A selected strand is identified from the shuffled nucleic acid strands for a criterion.
Claims21
A method comprising:
- a) fragmenting parent nucleic acids thereby generating three or more double-stranded nucleic acid fragments that have blunt ends;
- b) randomly ligating at least a subset of the blunt-ended double-stranded nucleic acid fragments to a hairpin nucleic acid and generating shuffled nucleic acids such that 5′ and 3′ ends of the shuffled nucleic acids are capped and can no longer ligate to other nucleic acid molecules;
- c) amplifying the shuffled nucleic acids using a primer that anneals to the hairpin nucleic acid thereby generating amplified shuffled nucleic acids;
- and
- d) identifying a first selected nucleic acid for a non-coding property from the amplified shuffled nucleic acids.
2 more elements
A method comprising:
- a) fragmenting a parent nucleic acid thereby generating three or more double-stranded nucleic acid fragments;
- b) modifying the double-stranded nucleic acid fragments such that the double-stranded nucleic acid fragments have blunt ends;
- c) ligating the blunt ends of at least a subset of the double-stranded nucleic acid fragments at random to a hairpin nucleic acid and generating shuffled nucleic acids such that 5′ and 3′ ends of the shuffled nucleic acids are capped and can no longer ligate to other nucleic acid molecules and at least one of the shuffled nucleic acids is non-homologous or in vitro non-homologous to the parent nucleic acid;
- d) amplifying the shuffled nucleic acids using a primer that anneals to the hairpin nucleic acid thereby generating amplified shuffled nucleic acids;
- and
- e) identifying a first selected nucleic acid for a non-coding property from the amplified shuffled nucleic acids.
3 more elements
Dependencies are inferred from the claim text.