cushion Protocols

Category: Cell Organelle Protocols: Nucleus Protocols

The key step is the lysis which solubilizes centrosomes away from nuclei by very low ionic strength lysis after treatment of cells with nocodazole and cytochalasin B. The released centrosomes are then centrifuged onto a Ficoll cushion (to avoid pelleting) and the interface between the lysate and the Ficoll is collected and the centrosomes are concentrated on a sucrose gradient. Fractions are assayed by spindown and double IF with 5051 serum and anti-tubulin and the pooled fractions are frozen... - [Read CHO Centrosome Prep Protocol]

Category: DNA Protocols: DNA Extraction Protocols: DNA Extraction Polyacrylamide Gels

DNA Electroelution.  This protocol describes the purification of DNA by trapping in a high-salt cushion in a "UEA AnalyticalElectroeluter" (IBI).  This machine is no longer manufactured, to our knowledge.  However, a smiliar device can be easily made from Plexiglas according to the following diagram, taken from Cornel Mulhardt, Molecular Biology and Genomics (2007) Academic Press, p.52: Schimenti Lab - [Read DNA Electroelution]