Experimental Metagenome on MinION

This is an artificial metagenome created by pooling various proportions of DNA from four microbe cultures to simulate a simple metagenome for analysis by Oxford nanopore MinION. Single-species cultures of Escherichia coli, Pseudomonas fluorescens, Microcystis aeruginosa, and Synechococcus elongatus were grown to log phase then harvested for DNA extraction. Following mechanical shearing, templates were prepared using either equimolar amounts of each species or combinations of DNAs where one species was rare (0.05%). Libraries were created using his-tag bead purification according to MinION specifications and libraries were sequenced using MinION R7.x flow cells. A separate library was created by pre-amplifying with Phi-29 a commercial mixture of DNAs in staggered proportions from 20 bacterial species.