METHOD FOR SELECTIVE AMPLIFICATION OF DNA FRANGMENTS FOR GENETIC FINGERPRINTING

摘要

The invention discloses a method for preparing a reproducible selective set of DNA fragments from the DNA or cDNA of one or more organisms as a main step in a DNA fingerprinting method. The method comprises a) cutting DNA with two or more restriction enzymes to obtain a set of double stranded DNA fragments, b) ligating to the DNA fragments double stranded adapters that match the generated fragment ends, and c) performing a polymerase chain reaction using primers which are specific for the adapters to obtain the reproducible selective set of DNA fragments. In step b) at least one modified adapter is used which is composed of two oligonucleotides that are only partly complementary, leaving at least one part of the adapter single stranded, in such a way that a complementary strand of said single stranded part of the adapter cannot be synthesized from an available 3'-end in the adapter itself, and in step c) the primer which is specific for the modified adapter, is identical or similar to the single stranded part of the adapter, such that only after complementation of the single stranded part of the adapter said primer can anneal to the complement, and in such a way that this primer can be extended in the direction of the ligation site.