Rolling circle amplification of circular genomes

摘要

Disclosed are compositions and a method for amplification of circular genomes. The method is based on rolling circle amplification of the circular genomes which involves strand displacement replication by primers. The disclosed method allows differential amplification of circular genomes of interest. In genomic nucleic acid samples containing both a circular genome of interest and non-target nucleic acids, such as non-target genomes, the disclose methods and compositions can result in many-fold differential amplification of the circular genome of interest over non-target nucleic acids. It has been discovered that selection of a set of primers complementary to a circular genome of interest can result in much greater amplification of the circular genome of interest relative to non-target nucleic acids present. Such differential amplification of circular genomes is very useful for obtaining useful amounts of genomes of interest from a mixed nucleic acid sample. For example, mitochondrial genomes, which, absent complicated and time consuming purification, are in the presence of non-target nucleic acids (such as the host cell genome), can be differentially amplified relative to the host cell genome and other non-target nucleic acids using the disclosed methods and composition.