Novel oligonucleotide arrays and their use for sorting, isolating, sequencing, and manipulating nucleic acids
The invention provides hyperactive mutant recombinases and hybrid mutant recombinases, and methods for their identification. Also provided are nucleic acids encoding hyperactive mutant recombinases and hybrid recombinases, as well as vectors and host cells. Host cells include eukaryotic cells capable of expressing said recombinases and carrying out site-specific recombination in the cell. The mutant recombinases may be used, for example, in biotechnology, gene therapy or transgenic applications.
1 - 159 . (canceled)
160 . A composition comprising at least two oligonucleotides complementary to unique sequences of interest in a DNA sample, and an amplified mixture of DNA isolated from a genome, wherein the amplified mixture of DNA is made by cleaving a genomic DNA sample with a restriction enzyme, thereby providing restriction fragments;
ligating linker nucleic acids to the DNA restriction fragments;
providing oligomers that are complementary to the linker nucleic acids; and,
amplifying the linked DNA by the polymerase chain reaction using the oligomers as primers, thereby providing the amplified mixture of DNA.
161 . The composition of claim 160 , wherein the at least two oligonucleotides are immobilized on a solid substrate.
162 . A method of characterizing a nucleic acid, comprising:
providing at least one oligonucleotide which is complementary to a unique sequence of interest in a genomic DNA sample;
amplifying a mixture of nucleic acids comprising a group of genome fragments, thereby providing an amplified nucleic acid mixture of genome fragments; and
hybridizing the at least one oligonucleotide to the amplified nucleic acid mixture, thereby detecting at least one nucleic acid of interest in said amplified mixture.
163 . The method of claim 162 , wherein the oligonucleotide is a member of an array of oligonucleotides, which array comprises additional oligonucleotides which hybridize to unique genomic sequences of interest.
164 . The method of claim 162 , wherein the amplified nucleic acid mixture is made by cleaving a genomic DNA sample with a restriction enzyme, thereby providing restriction fragments;
ligating linker nucleic acids to the DNA restriction fragments;
providing oligomers that are complementary to the linker nucleic acids; and,
amplifying the linked DNA by the polymerase chain reaction using the oligomers as primers, thereby providing the amplified nucleic acid mixture.