METHOD FOR AMT-RFLP DNA FINGERPRINTING

1 . A method for identifying a sample organism, comprising the steps of:

obtaining a sample of DNA;

isolating a portion of said sample to be analyzed;

cutting said portion into at least two fragments;

ligating both ends of said fragments with double-stranded linker sequences;

adding said fragments to polymerase chain reaction using a dye; and

analyzing said fragments using a melt-curve analysis.

2 . The method of claim 1 wherein said sample organism is selected from a group consisting of bacterial, fungal, viral, plant, animal, and combinations thereof.

3 . The method of claim 1 wherein said dye is selected from the group consisting of SYBR green-intercalating dyes, cyanine dimer dyes, ethidium bromide dyes, and acridine dyes.

4 . The method of claim 1 wherein said cutting step further comprises enzymatically cutting said sample by using a restriction endonuclease that recognizes a specific DNA cut site.

5 . The method of claim 4 wherein said isolation step further comprises treating said fragment with a phosphatase that removes phosphates from each end of said fragment.

6 . The method of claim 1 wherein said adding step further comprises using at least one said fragment as a template.

7 . The method of claim 4 further comprising the step of:

amplifying said fragments using one primer complementary to said ligated linker sequence and one primer complementary to a universal DNA sequence, wherein amplification produces at least two new strands.

8 . The method of claim 1 wherein said analyzing step further comprises:

denaturing said fragments at a high temperature;

lowering said temperature until said fragments reassemble;

depositing said dye between said fragments as said fragments are reassembling;

increasing said temperature until said fragments begin to break apart;

monitoring said dye and said fragments as said temperature is increased;

recording at least three data points; and

analyzing said data points with known information to identify said sample.

9 . The method of claim 1 wherein said sample has a length of from about 0.1 nanograms to 1000 nanograms.

10 . The method of claim 1 wherein said method is automated.

11 . A method for identifying a sample organism, comprising the steps of:

obtaining a sample of DNA;

cutting said sample by using a restriction endonuclease that recognizes a specific DNA cut site, wherein cutting produces at least one fragment of said sample;

treating said cut sample with a phosphatase to remove phosphates from ends of said fragments;

ligating a double-stranded linker sequence to each end of said fragments;

adding at least one said fragment to a polymerase chain reaction using a SYBR green-intercalating dye, wherein said fragment is used as a template;

amplifying said fragments by using one primer complementary to said ligated linker sequence and one primer complementary to a universal DNA sequence, wherein amplification produces at least two strands;

denaturing at least two said strands at a high temperature;

lowering said temperature until said strands reassemble, wherein said SYBR green dye is deposited between said strands;

recording at least three data points as said temperature is slowly increased;

comparing said data points with known information about organisms; and

determining said identification of said sample organism.