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Yeast cells had been grown instantly from 30��C throughout SDC-leu medium and gathered with an OD600 associated with 3. Cells had been lysed inside 25?ml cold barrier 1 (20?mM Tris-HCl pH 7.5, 150?mM NaCl, 50?mM K(OAc), 2?mM Milligram(OAc)2) formulated using protease chemical combination FY (Serva), 1?mM phenylmethylsulfonyl fluoride (PMSF), 1?mM dithiothreitol (DTT), and also 0.1% (v/v) NP-40 (ly

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