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[Cell Culture] In Vitro Transcription with SP6 Polymerase 日期：2006-07-24 14:11:15 点击：19 好评：0
Monday, October 20, 2003 Description All reagents and tubes should be RNase-free. Make solutions up in plastic vessels using ddH2O. Autoclave the NaCl, MgCl2, Tris, glycerol, and gelatin solutions. Do not touch the undersides of tube c...
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[Cell Culture] Labeling Oligonucleotides 日期：2006-07-24 14:11:14 点击：25 好评：0
Tuesday, October 21, 2003 Description Labeling Oligonucleotides Procedure 1. Combine the following reactants in a microcentrifuge tube: 0.5 靗 of Denaturing Buffer 50 ng of the Oligonucleotide to be labeled (in TE Buffer) Bring the total r...
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[Cell Culture] Chemotaxis Assay Using Microchemotaxis Chambers (Modified Bo 日期：2006-07-24 14:11:14 点击：25 好评：0
Tuesday, October 21, 2003 Description This assay uses a simple apparatus to study chemotaxis of leukocytes or other migratory cells. The apparatus consists of two multi-well chambers separated by a filter containing pores of uniform size....
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[Cell Culture] Human Peripheral Blood Mononuclear Cell Preparation 日期：2006-07-24 14:11:13 点击：34 好评：0
Tuesday, October 21, 2003 Description This protocol describes a procedure for isolating human peripheral blood mononuclear cells (lymphocytes and monocytes) from a Buffy Coat (obtained from a blood bank). Platelets, red blood cells, and ne...
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[Cell Culture] TransWell Chemotaxis 日期：2006-07-24 14:11:12 点击：36 好评：0
Tuesday, October 21, 2003 Description This protocol describes a method for studying chemotaxis (migration towards a concentration gradient of chemoattractant) of leukocytes (neutrophils, monocytes and lymphocytes) or other migratory cells....
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[Cell Culture] G1/S Phase Block for Cell Synchronization 日期：2006-07-24 14:11:12 点击：36 好评：0
Monday, October 27, 2003 Description G1/S Phase Block for Cell Synchronization Procedure 1. Presynchronize cells at G0 by serum starvation. Add Low Serum Medium to cells at 70% confluency and allow cells to grow in this medium for 2 days....
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[Cell Culture] G2 Phase Block with Hoechst 33342 for Cell Synchronization 日期：2006-07-24 14:11:11 点击：50 好评：0
Monday, October 27, 2003 Description G2 Phase Block with Hoechst 33342 for Cell Synchronization Procedure 1. Presynchronize cells at G0 by serum starvation. Add Low Serum Medium to cells at 70% confluency and allow cells to grow in this me...
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[Cell Culture] Metaphase Block for Cell Synchronization 日期：2006-07-24 14:11:11 点击：36 好评：0
Monday, October 27, 2003 Description Metaphase Block for Cell Synchronization Procedure 1. Remove the medium from an exponentially-growing cell culture and rinse it with 10 ml of PBS. (See Hint #1) 2. Replace the PBS with Complete Medium w...
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[Cell Culture] Preparing Chromosome Spreads 日期：2006-07-24 14:11:10 点击：18 好评：0
Monday, October 27, 2003 Description Preparing Chromosome Spreads Procedure 1. Expose an exponentially-growing culture of cells to Colcemid (CAUTION! see Hint #1) in culture medium to arrest cells in the mitotic phase of the cell cycle (se...
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[Cell Culture] MITOCHONDRIAL DNA ISOLATION 日期：2006-07-24 14:11:10 点击：20 好评：0
Friday, November 07, 2003 Description MITOCHONDRIAL DNA ISOLATION Procedure Grind in mortar and pestle or Waring blender with 5-7 volumes buffer A per g tissue. Use MCE at 350 l/L, and if necessary, with 5 ml 1 M DIECA/L. Squeeze through c...