Salmon sperm dna ssdna

AL‐I‐OH and AL‐II‐NOH were synthesized in our laboratory as described (Attaluri et al. 2014), and AA‐I was purified from A. indica by high‐performance liquid chromatography. AA‐related compounds were dissolved in dimethyl sulfoxide (DMSO) at 30–40 mM and stored at −20°C. 2,6‐Dichloro‐4‐nitrophenol (DCNP) was purchased from Tokyo Chemical Industry Co. LTD (Japan). Enzymes used for ³²P‐postlabeling analysis were obtained from Worthington (Newark, NJ), New England Biolabs (Ipswich, MA), MP Biomedicals (Solon, OH), and Sigma Aldrich (St. Louis, MO). DMSO, 3′‐phosphoadenosine‐5′‐phosphosulfate (PAPS) (70% purity), salmon sperm DNA (ssDNA), pentachlorophenol (PCP), quercetin, β‐estradiol, 4‐nitrophenol were from Sigma‐Aldrich. PAPS (>90% purity) and 3′‐phosphoadenosine‐5′‐phosphate (PAP) were purchased from R&D Systems (Minneapolis, MN). dG‐AL‐II and dA‐AL‐II containing oligonucleotides were synthesized as described (Attaluri et al. 2010). γ‐³²P‐ATP (6000 μCi/mmol) was obtained from PerkinElmer (Boston, MA). Recombinant human SULT1B1 was purchased from MyBioSource Inc. (San Diego, CA).

Salmon Sperm DNA (ssDNA) (Sigma-Aldrich) with a concentration of 100 μg/mL was incubated with different concentrations of Complex 1 for 30 min in a 37 °C incubator. Then, the mixture was added to the C. neoformans culture and incubated at 37 °C for 24 h in tubes with constant shaking. The number of viable cells was determined by plating the suspension on drug-free solid media and then measuring CFUs.

All osmolytes (sucrose, mannose, sorbitol, mannitol, trimethylamine N-oxide, trehalose), amino acids, lithium acetate, Tris-HCl, EDTA and salmon-sperm DNA (ssDNA) were purchased from Sigma-Aldrich (St. Louis, MO). Yeast extract, dextrose, peptone, agarose and polyethylene glycol (PEG) were purchased from Hi-media Laboratories (Mumbai, India).