Alu PCR: The Use of Repeat Sequence Primers for Amplification of Human DNA from Complex Sources

Abstract

The polymerase chain reaction (PCR) has revolutionized the isolation and analysis of specific nucleic acid fragments from a wide variety of sources.¹ However, application of the PCR to isolate and analyze a particular DNA region has required knowledge of DNA sequences flanking the region of interest. This limits amplification to regions of known DNA sequence. We sought to amplify human DNA of unknown sequence from complex mixtures of human and other species DNAs. In particular, we have applied the PCR to isolation of human DNA specifically from somatic cell hybrids retaining human chromosome fragments in rodent cell backgrounds. This allows the isolation and characterization of sequences from specific human regions retained in hybrids, obviating the requirement of cloned DNA libraries and isolation of human clones through the use of human-specific repeat sequence probes.² The method, Alu PCR, has also proven useful for the rapid isolation of human insert DNA from cloned sources as well, extending the application of the PCR to genomic DNAs cloned in lambda and yeast artificial chromosome (YAC)³ vectors. The adaptation of the PCR to large genomic regions provides another tool for the analysis of the human genome.