Drummond:Protocols

Local Protocols

Here are some protocols we use in the lab.

• Chloroform/methanol precipitation of proteins
• Resolving and detecting ubiquitin
• Using paromomycin to alter translation in yeast
• Cell preparation and protocol to view RFP/TFP fluorescence
• Using the Zeiss LSM 510 confocal microscope
• Yeast OD and cell viability model
• Standard conditions for flow cytometer growth rate assay
• High-efficiency yeast transformation
• Yeast genomic DNA prep (consider the rather rapid yeast genomic DNA prep instead)
• Isolating total, soluble and insoluble protein fractions
• Making chemically competent E. coli cells
• Preparing anti-ubiquitin beads
• Periplasmic protein prep from E. coli

Remote Protocols

• Spore enrichment for random-spore analysis (from Rockmill Meth. Enz. 1991)
• Rather rapid yeast genomic DNA prep (Gottschling Lab)
• SDS-PAGE (Szostak Lab)

Data Analysis Protocols

• Akashi's test for translational accuracy selection

Notes

• The relationship between the selection coefficient and relative growth rates: under construction
• Thoughts on lysis of yeast cells