Methods for forming recombined nucleic acids

Abstract

Described herein are methods for generating recombined nucleic acids. In one method, fragments of a sequence are provided wherein the fragments have non-extendable 3′ ends. A primer is provided and the primer and fragments are reacted under conditions to extend the primer to form a recombined nucleic acid molecule. In the methods herein, the non-extendable fragments act only as templates, rather than as templates and primers.

1 - 14 . (canceled)  
   
   
       15 . A vector comprising a recombined nucleic acid molecule, wherein said recombined nucleic acid molecule is formed by: a) randomly fragmenting at least one initial double-stranded nucleic acid molecule to produce a plurality of fragments; b) adding a terminator molecule to the 3′ ends of said fragments to provide a plurality of double-stranded template fragments; c) denaturing the strands of said double-stranded template fragments to produce single-stranded template fragments; d) annealing at least one primer to said single-stranded template fragments under conditions such that said primer produces a complementary strand of said template fragments; and e) repeating steps c) and d) to produce a recombined nucleic acid molecule, wherein said recombined nucleic acid molecule is approximately the same length as the initial double-stranded nucleic acid molecule.  
   
   
       16 . A cell comprising the vector of  claim 15 .  
   
   
       17 . A pool of recombined nucleic acid molecules generated by the method of: a) randomly fragmenting at least one initial double-stranded nucleic acid molecule to produce a plurality of fragments; b) adding a terminator molecule to the 3′ ends of said fragments to provide a plurality of double-stranded template fragments; c) denaturing the strands of said double-stranded template fragments to produce single-stranded template fragments; d) annealing at least one primer to said single-stranded template fragments under conditions such that said primer produces a complementary strand of said template fragments; and e) repeating steps c) and d) to produce a recombined nucleic acid molecule, wherein said recombined nucleic acid molecule is approximately the same length as the initial double-stranded nucleic acid molecule.  
   
   
       18 . A recombined nucleic acid molecule formed by the method of: a) randomly fragmenting at least one initial double-stranded nucleic acid molecule to produce a plurality of fragments; b) adding a terminator molecule to the 3′ ends of said fragments to provide a plurality of double-stranded template fragments; c) denaturing the strands of said double-stranded template fragments to produce single-stranded template fragments; d) annealing at least one primer to said single-stranded template fragments under conditions such that said primer produces a complementary strand of said template fragments; and e) repeating steps c) and d) to produce a recombined nucleic acid molecule, wherein said recombined nucleic acid molecule is approximately the same length as the initial double-stranded nucleic acid molecule.