Welcome to Heyer Lab Protocoals

My thanks to everybody, who contributed.

Genreral information

Saccharomyces cerevisiae

MMS Handling

Some useful tips ... (from PLOS Computational Biology)

1. Ten Simple Rules for a Good Poster Presentation
2. Ten Simple Rules for Getting Published
3. Ten Simple Rules for Making Good Oral Presentations
4. Ten Simple Rules for Getting Grants
5. Ten Simple Rules for a Successful Collaboration

Current Protocols

Media/ Ampicillin stock

How to archive strains?

E. coli:

• transformation
• quick extract for protein analysis
• plasmid miniprep
• purification of his-tagged proteins

Yeast:

• transformation
• KanMX disruption with PCR
• chromosomal DNA preparation (Cao/Lichten)
• plasmid retrieval
• galactose induction and protein extract
• mating, sporulation and tetrad analysis
• IR irradiation
• spotting assay for growth

DNA/RNA:

• plasmid restriction and agarose gel
• polyacrylamide gel
• ligation (sticky ends, blunt ends)
• kinase treatment
• DNA extraction with CTAB
• crosslinking with psoralen (DNA prep, new X-linking protocol, reversal)
• CHEF gels (agarose plugs, running)
• 2d gel
• southern blot
• PCR
• ethanol precipitation
• phenol extraction

Protein:

• SDS-PAGE and coomassie staining
• western blot
• immunoprecipitation (IP)
• GST pulldown
• ATPase assay
• DNA strand exchange assay
• antibody production
• kinase assay
• nucleoprotein gel assay
• boiling dialysis tubes
• TCA precipitation

Yeast GFP Construct Database:

• From SGD
  
• From the University of Washington