Molecular markers for dna

Escherichia coli strain TOP10 was used as host for plasmid cloning experiments. Bacteria were grown in Luria–Bertani medium supplemented with ampicillin (100 mg mL⁻¹) at 37°C. Saccharomyces cerevisiae Wa6 (ade, his7-2 leu2-3 leu2-112 ura3-52) the host for protein expression studies. Recombinant yeast was grown in S medium (7 g L⁻¹ yeast nitrogen base, 1 g L⁻¹ casamino acids, 20 g L⁻¹ glucose supplemented with 50 mg mL⁻¹ histidine, 200 mg mL⁻¹ adenine, and 50 mg mL⁻¹ leucine) for 2 days with shaking at 30°C. Culture media components and agarose were purchased from Sigma-Aldrich, USA and liquid-chromatography grade solvents and buffer components were supplied by Merck, Germany. Isopropyl-b-D-thiogalactopyranoside (IPTG), X-Gal, restriction enzymes, RNase A, molecular markers for DNA, and dNTPs were purchased from Fermentas (Vilnius, Lithuania). Oligonucleotides, aniline, thiobenzamide, cumene hydroperoxide, fatty acids, antibiotics, and amino acids were supplied by Sigma-Aldrich, USA.