ale yeast 454 reads
To minimize the amount of mitochondrial DNA during sequencing, GSY2239 was plated on YEP-glucose containing 30 ug/ml of ethidium bromide, a protocol that is known to induce petite mutations. After two days, multiple colonies were isolated and lack of growth on a non-fermentable substrate (YEP-glycerol) was confirmed. For genomic DNA extraction, a single colony, GSY2239.1 was grown overnight in 3 ml YEP + 2% dextrose. Cells were resuspended in 200 ul Triton X-100, 1% SDS, 100 mM NaCL, 50 mM Tris-Cl (pH 8.0) and 1 mM EDTA (pH 8.0) whereby DNA was extracted using phenol:chloroform and resuspended in 200 ul TE RNAse (50 ug/ml).