Sunday, October 19, 2003 Description Crosslinking DNA-Binding Protein to DNA Using a Bromodeoxyuridine-Substituted Probe Procedure 1. Combine 10 g M13 ssDNA vector and an equal molar amount of 17 bp M13 universal primer in a microcentrifuge...

Saturday, October 18, 2003 Description Cells that were previously transfected with a lac Z construct can be stained for -Galactosidase activity. Procedure 1. Wash the cells 2 to 3 times in 2 ml of PBS solution (see Hint #2 and Hint #3). 2....

Saturday, October 18, 2003 Description This protocol uses a lithium acetate/PEG treatment to transform yeast with DNA. Plasmid DNA or linear DNA may be used Procedure 1. Use cells from an overnight yeast culture to inoculate 300 ml of liqu...

Thursday, October 16, 2003 Description A method that worked for me, pretty close to the commercial description Procedure 1 - Preparation of cells for transfection Day 1: Plate the cells one-day before the transfection experiment. The appro...

Thursday, October 16, 2003 Description Protocol for large scale Plasmid DNA isolation Procedure Y To be read as micro (mu) 1.5ml of LB medium containing 50Yg/Yl ampicillin [Sigma A-0166] was innoculated with a single bacterial colony. The...

Wednesday, October 08, 2003 Description For preparation of competent bacterial cells (14), a glycerol cell culture stock of the respective E. coli strain is thawed and added to 50 ml of liquid media. This culture then is preincubated at 37...

Wednesday, October 08, 2003 Description DNA ligations are performed by incubating DNA fragments with appropriately linearized cloning vector in the presence of buffer, rATP, and T4 DNA ligase (1,2). For random shotgun cloning, sonicated or...