The tobacco genome is estimated to be approximately 4.5Gb in size

GS FLX Titanium data representing approximately 17× depth of the tobacco genome was assembled using gsAssembler v2.8 (Roche454, Branford, CT) using default settings with the exception of minimum identity and minimum overlap length being set to 97% and 100 bp respectively. This assembly was corrected for sequence errors, including homopolymers, by mapping the 300 bp Illumina PE sequence to the assembly, calling the SNPs, filtering them based on the read coverage and the allele frequency and applying the Perl script Vcf4FastaCorrection (available from https://github.com/aubombarely/GenoToolBox/blob/master/SeqTools/Vcf4FastaCorrection). The resulting WGS assembly contained 440,772 contigs/scaffolds and covered 4,145,428,719 bp (3,510,246,061 bp of which were defined), with an N50 of 334,966 bp.