Method for designing probe in DNA microarray, and DNA microarray provided with probe designed thereby
Provided is a probe to be used in a DNA microarray having an excellent detection rate of a polymorphism such as SNP contained in genomic DNA. A method for designing a probe according to the invention includes the steps of: specifying one or more regions covering at least a part of fragments flanked by restriction enzyme recognition sites recognized by a restriction enzyme, contained in genomic DNA derived from an organism to be tested; and designing a probe for the specified one or more regions for detecting the fragment in the organism to be tested.
1. A method for detecting a mutation using a DNA microarray having a plurality of polynucleotide probes immobilized thereon, said method comprising the steps of:
extracting a genomic DNA derived from an organism to be tested;
digesting the genomic DNA with a restriction enzyme having the same recognition sequence as a restriction enzyme used to design the probes immobilized on the DNA microarray;
connecting an adaptor to the genomic DNA fragments obtained by the restriction enzyme treatment;
amplifying the genomic DNA fragments using a primer capable of hybridizing to the adaptor; and
detecting a hybrid of a genomic DNA fragment containing a mutation with a probe capable of detecting the mutation, by bringing the amplified genomic DNA fragment into contact with the DNA microarray,
wherein said DNA microarray comprises a plurality of probes, and a carrier on which the probes are immobilized, wherein said plurality of probes is capable of detecting a mutation contained in a genomic DNA fragment obtained by said digesting, wherein the plurality of polynucleotide probes comprises probes covering different portions of the genomic DNA fragment, wherein the plurality of polynucleotide probes covers the entire region of the genomic DNA fragment, and wherein the probes in said plurality of polynucleotide probes are shorter in length than the genomic DNA fragment.
2. The method according to claim 1 , wherein, in the step of digesting the genomic DNA, the genomic DNA is digested with a plurality of restriction enzymes.
3. The method according to claim 2 , wherein, in the connecting step, an adaptor is connected which corresponds to one restriction enzyme from said plurality of restriction enzymes, or which corresponds to several of the restriction enzymes in said plurality of restriction enzymes.
4. The method according to claim 1 , wherein the adaptor has a complementary sequence to a protruding end of the genomic DNA fragments obtained in the step of digesting the genomic DNA with a restriction enzyme.
5. The method according to claim 1 , wherein the organism to be tested is different from the organism used in preparing the DNA microarray.