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[Enzymology] Sephadex G-50 spun column purification 日期：2005-07-18 00:00:00 点击：347 好评：0
Wednesday, November 19, 2003 Description Spin column purification can be used to change buffers without a concomitant change in solution volume, to remove protein contaminants or to purify plasmid DNA suitable for sequencing. Procedure 1)...
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[Enzymology] Yeast Nuclear Extract 日期：2005-07-18 00:00:00 点击：179 好评：0
Tuesday, November 18, 2003 Description Yeast Nuclear Extract Procedure Cell Growth 3 liters of cells are grown in YPD (3% glucose) to A600 of 3-5. Antifoam A (two drops from a P200 pipetman) is added to media before autoclaving. For wild t...
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[Enzymology] Yeast Sucrose Gradient Protocol 日期：2005-07-18 00:00:00 点击：175 好评：0
Tuesday, November 18, 2003 Description Yeast Sucrose Gradient Protocol Procedure Grow a 2 ml culture, 24 hr. at 30oC in selective media When culture is ready, use it to inoculate about 55 ml (50 ml plus 5 for O.D.600 readings) of selective...
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[Enzymology] Yeast Glass Bead Prep 日期：2005-07-18 00:00:00 点击：261 好评：0
Tuesday, November 18, 2003 Description Yeast Glass Bead Prep Procedure Sample Buffer: 0.06M Tris-HCl, pH 6.8 10% (v/v) glycerol 2% (w/v) SDS 5% (v/v) 2-mercaptoethanol 0.0025% (w/v) bromophenol blue 0.1 M PMSF 0.5 M Benzamidine 1. Grow 25...
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[Enzymology] Yeast Rapid Protein Prep 日期：2005-07-18 00:00:00 点击：192 好评：0
Tuesday, November 18, 2003 Description Yeast Rapid Protein Prep Procedure Sample Buffer: 0.06M Tris-HCl, pH 6.8 10% (v/v) glycerol 2% (w/v) SDS 5% (v/v) 2-mercaptoethanol 0.0025% (w/v) bromophenol blue Grow cells overnight (~1E7 cells/ml)...
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[Enzymology] Yeast Cell Lysates 日期：2005-07-18 00:00:00 点击：223 好评：0
Tuesday, November 18, 2003 Description Yeast Cell Lysates Procedure Grow cells to 0.3 to 0.5 O.D.(600) (~1E7 cells/ml). Use about 1.0 to 5.0 OD of cells per lysate; if your cells are at 0.5 OD per ml, use 2 ml to give you 1.0 OD of cells i...
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[Enzymology] Yeast Whole Cell Extracts 日期：2005-07-18 00:00:00 点击：170 好评：0
Tuesday, November 18, 2003 Description Yeast Whole Cell Extracts Procedure -Use sterile technique and sterile solutions in steps 1 to 3.- 1. Using a saturated starter culture, inoculate 25 to 30 ml of appropriate media in a 125 ml flask. *...
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[Enzymology] MAKING YEAST EXTRACTS for WESTERNS 日期：2005-07-18 00:00:00 点击：233 好评：0
Tuesday, November 18, 2003 Description MAKING YEAST EXTRACTS for WESTERNS Procedure Harvest 5ml cells at OD = 1.0 in 15ml falcon tube[Optional : freeze pellet in liquid nitrogen] r/s in 500ul cold TCA buffer w/ 2X Protease inhibitors prepa...
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[Enzymology] Actin Capture Assay 日期：2005-07-18 00:00:00 点击：255 好评：0
Tuesday, November 18, 2003 Description Actin Capture Assay Procedure Dialyze purified GST fusion proteins and actin into PBS + 1mM MgCl2 . Mix 5ug actin into 50ul total volume binding buffer. Mix 5ug GST-fusion protein into total volume 50...
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[Enzymology] Purification of the NSP1 RD Protein 日期：2005-07-18 00:00:00 点击：162 好评：0
Tuesday, November 18, 2003 Description Purification of the NSP1 RD Protein Procedure Day 1 1. Set up an overnight flask of 2 litres 2XTY Amp in a 2 litre flask, shake fairly gently at 37C, or more vigorously at 34C. 2. Harvest cells, resus...