Arbitrarily primed PCR for comparison of meta genomes and extracting useful loci from them

Abstract

A method is described that uses arbitrarily primed PCR followed by many cycles of amplification under stringent conditions and selection by computational means to obtain a set of sequence tags that can be used for the comparison of metagenomes. Relative to unselective shot-gun sequencing, the results are small data sets that can be csompared electronically or plotted as scattergrams that are simple to interpret. The method can be used to compare groups of samples of any size to build in-house databases from which, for example, the provenance of trace soil samples may be inferred. The method also allows for selection of primers with locus-specificity and an example is given in which a South Australian sequence-related to a Portuguese thermophile (Rubrobacter radiotolerans) is extracted and tested on a set of soils.