The Building Blocks, Biological Pathways and Networks program has transitioned from Common Fund support.  For more information, please visit https://commonfund.nih.gov/bbpn/index. Please note that since the Building Blocks, Biological Pathways and Networks program is no longer supported by the Common Fund, the program website is being maintained as an archive and will not be updated on a regular basis. 

BOEKE Protocols

• Yeast high efficiency transformation - dSLAM (PDF) 
• Yeast sporulation - dSLAM (PDF) 

COLE Protocols

• Chemical synthesis of a novel coenzyme A-based affinity probe (PDF) 

COTTER Protocols

• Quantification of lysine acetylation (PDF) 
• N-terminal derivatization for de novo sequencing (PDF) 
• Determination of ubiquitylation sites (PDF) 
• Determination of SUMOylation sites (PDF) 

Frommer Protocols

• Protocol for monitoring intracellular metabolite levels

MBIC Protocals

• Preparing stock solutions of Malachite Green derivatives 

MELUH Protocols

• Indirect Immunofluorescence for Yeast (PDF) 
• DAPI Staining Yeast (PDF) 
• Chromatin Immunoprecipitation (ChIP) for Yeast (PDF)

PANDEY Protocols

• 18O - labeling trypsin digest protocol for relative quantitation by mass spectrometry 

RANDO Protocols

• Mononucleosome Chromatin Immunoprecipitation for Yeast (PDF) 

NCDIR Protocols

Disruption of Yeast or Bacteria Cells

• Harvesting Cells and Making Noodles
• Cryogenic Disruption Using PM100 Ball Mill
• Microfluidizer

Subcellular Fractionation

• Yeast NE & NPC Preps
• Yeast Nuclei & SPB Preps
• Carbonate Extraction of NEs
• Rapid Subcell Fractionatn for Nucl & NEs
• Heparin Extraction of NEs

Isolation of Protein Complexes

• Conjugation of Tosylactivated Dynabeads
• Conjugation of Epoxyactivated Dynabeads
• Pullouts with IgG Resin
• Immunoaffinity purifications of protein complexes

Mass Spectrometry

• In-gel Digestion protocol 

Genomic Analyses

• mRNA expression microarrays

Protein Biochemistry

• Carbonate Silver Stain
• Electropurification
• Methanol Precipitation of Protein
• Protease Mapping
• Strip Blots
• Whole Cell Lysis and TCA Precipitation