MinION sequencing
The gDNA was sheared using g-TUBE spun at 2,000 x g for 2 min (Covaris, Woburn, Massachusetts).  DNA was pretreated with three NEB reagents in this order: PreCR Repair Mix, NEBNext End Repair Module, NEBNext dA-Tailing Module.  We then used the MinION MAP Standard Protocol for the gDNA Library Preparation Kit v004 (Oxford Nanopore Technologies, Oxford, UK) with no changes in the recommended steps.  The same library was run on two different flow cells.
Virginia Commonwealth University
ena-EXPERIMENT-Virginia Commonwealth University-05-03-2015-14:24:31:055-2
kit
SQK-MAP003
sequencing protocol
gDNA
dna extraction protocol
Single-species cultures of Escherichia coli, Paracoccus denitrificans, Microcystis aeruginosa, and Synechococcus elongatus were grown to log phase then harvested and extracted using the MoBio PowerSoil Kit (Carlsbad, CA).  DNA in 10 mM Tris-HCl was quantified and the four preparations were subsequently combined at 0.5 ug each for a total of 2 ug as the initial template for library preparation.