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...

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...

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...

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...

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....

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....

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...

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....

Tuesday, October 21, 2003 Description Labeling Oligonucleotides Procedure 1. Combine the following reactants in a microcentrifuge tube: 0.5 l of Denaturing Buffer 50 ng of the Oligonucleotide to be labeled (in TE Buffer) Bring the total re...

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 caps or pipette tips with...