Method for amplifying all of the DNA fragments in a sample, including small and damaged fragments, by pretreating the fragments with P1 nuclease

摘要

A method of amplifying all of the DNA fragments of a sample, including fragments that are small or damaged, comprises an initial step of generating free 3'-OH end with P1 nuclease. The method also provides for extraction of the DNA fragments from a sample and reduction of their size to between 100 and 300 base pairs. A poly(dX) oligonucleotide is added to the 3' ends of the DNA fragments using a terminal transferase, and the DNA is denatured and hybridized to a poly(dY) primer complementary to the poly(dX) ends. Finally, the DNA is polymerized with DNA polymerase. The resulting DNA can be amplified by denaturing and slowly renaturing the complementary strands.