forces Protocols

Category: Yeast Protocols: Yeast Genetics Protocols: YACs Protocols

Protocol describes methods for isolation of DNA from a strain of S. cerevisiae carrying a recombinant YAC. Because the linear YAC DNAs are sensitive to shearing forces, pipettes with wide-bore tips should be used to transfer DNAs. The method is suitable for preparing DNA that will be used for agarose gel electrophoresis, Southern blotting, subcloning, genomic library construction, PCR, or other methods that do not require intact high-molecular-weight DNA. - [Read Growth of S. cerevisiae and Preparation of DNA Protocol]

Category: Cell Biology and Cell Culture: Cell Adhesion Protocols

Many proteins and molecules promote cell adhesion including several cell surface carbohydrate binding proteins. Cell adhesion measurements on 96-well microtiter plate format are difficult due to the shear forces generated by washing the wells. The protocol here introduces the use of a liquid-filled wash chamber that separates unbound cells by gravity. This eliminates uncontrolled shear forces and passage of adherent cells through a liquid/air interface. John L. Magnani~GlycoTech Corporation. - [Read Measurement of Cell Adhesion Under Static Conditions]

Category: Cell Biology and Cell Culture

This protocol introduces the use of a liquid-filled wash chamber that separates unbound cells by gravity thereby eliminating uncontrolled shear forces and passage of adherent cells through a liquid/air interface. The cells are loaded with a fluorescent dye (6-carboxyfluorescein diacetate) for detection although other methods such as radioactive labels malabels may be used. This protocol is also useful for assaying molecules that promote or inhibit cell adhesion. - [Read Protocol for Measurement of Cell Adhesion Under Static Conditions]

Category: Viral Manipulation Protocols: Phage Protocols

Protocol describes methods for recovery and purification of recombinant clones of bacteriophage P1 or PAC DNAs from bacteria. Because of their large size, these DNAs are sensitive to shearing forces and must be handled carefully. This protocol generally yields P1 DNA that works well as a substrate or template in enzymatic reactions. - [Read Working with Bacteriophage P1 and Its Cloning Systems Protocol]