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文献和实验Gel-purified, agarased YACs for yeast transformation
109 cells per ml (2 X 109 cells per 0.5 ml plug). See plug-making protocol . 2. SeaPlaque GTG agarose (FMC, Rockland, ME, Cat. No. 50112) 3. Modified agarase buffer (10 mM BisTris-HCl, pH 6.5, 1mM EDTA, 100 mM NaCl). Make 10X concentrated
Preparation of Yeast DNA Embedded in Agarose Plugs
for a saturated culture. A 5 ml saturated culture will yield one plug at approximately 5 x 108 cells per plug. (Each plug holds slightly less than 0.5 ml). 2. Harvest the cells by centrifugation at 1300 x g for 5 min. Wash the cell
Preparation of Yeast DNA Embedded in Agarose Plugs
will yield one plug at approximately 5 X 108 cells per plug. (Each plug holds slightly less than 0.5ml). 2. Harvest the cells by centrifugation at 1300 x g for 5 min. Wash the cell pellet twice with 50mM EDTA, spinning 5 min at 1300 x g as above.
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