Yeast

• Chromatin IP's (Meluh)

• Chromatin IP's (Megee)

• Chromatin IP's (Mechanical Lysis)

• Flow Cytometry for Yeast: Cell Preparation

• FACS machine instructions

• Arresting Yeast

• High Efficiency Yeast Transformation (Gietz Long Protocol)

• Quick Yeast Tranformation (Gietz Short Protocol)

• Yeast Gene Knockouts

• Met15

• Plasmid Isolation from Yeast

• Sporulation

• Frogging

• Frozen Permanents

• Sending Strains

• YAC Transfer by KAR1

Microscopy

• DAPI Staining of Yeast Cells

• FISH for Yeast

• Quick Fixation of Yeast Cells for GFP Fluorescence

• Indirect Immunofluorescence for Yeast I

• Indirect Immunofluorescence for Yeast II: modifications for GFP fix.

• Rapid DAPI Screen For Chromosome Missegregation Mutants

• Pds1:HA Immunofluorescence

• Yeast Chromatin Spreads

• Yeast Chromatin Spreads II (Guacci)

• Meiotic Spreads

• Meiotic Synchrony
  

DNA/RNA

• Best PCR

• DNA Fragment Purification: Low Melting Agarose

• DNA Fragment Purification: Glass Wool

• Isolation of High MW Yeast DNA

• Posiblot

• SDS Hybridization Solutions for Southerns

• Yeast RNA Isolation

• Northern Blots

• Pulse Field Electrophoresis

Protein

• Yeast Protein Prep (No Boil)

• Yeast Protein Prep (Boil)

• SDS Page (Biorad)

• Western

• Western Blot with BSA

• Carbonate Solution For Western Blot

• Silver Stain

Reagents and Equipment

• Lab Antibodies: Conc. for Western & IF

• ChIP Primers for Yeast Chromosomes

• Alpha Factor

• Ampicillin

• Oxalyticase

• Prep of Protein A Sepharose Beads

• Prep of Glass Beads for Yeast Lysis

• RNAse

• Salmon Sperm DNA

• SCE

• Spectrophotometers: Instructions for Lab and Upstairs

• Solutions for Posiblot transfer: neutralization and denaturing

• Zymolyase

• 10X TBE

• 20X SSPE

• EDTA

Media

• YPD Plates

• YPD Media (1L)

• YP Media (Large Batch)

• YPG Plates

• 2X N

• 2X YP Media

• Adenine Stock

• 2X Agar for bacteria and yeast

• B plates

• Supplements

• 20% Dextrose

• 30% glycerol

• Dropout Plates (500 ml)

• Dropout Plates (3L)

• 0.3% KOAc

• L-Broth

• L-Broth + Ampicillin

• L-Plates

• M9 Pro Thr Plates (Bacterial Media for Growing pDK plasmids)

• Benomyl Plates

• 5-FOA Plates

• G418 YEP plates

• G418 Dropout plates

• Clonat Plates

• Lead Plates

Computer

• Making PDF files

• Making and Ordering Oligos

• Quantitation of EtBr Gels (Image Quant & Excel)

Bacteria

• Frozen competent E. coli cells

• Electroporating E. coli

• Site-directed mutagenesis with oligos

• STET preps for rapid recovery of plasmid DNA

• Frozen Permanents

• Sending Strains

Microtubules

• Minichromosome-Microtubule Binding Assay

• Making Microtubule Seeds

• Cycling Tubulin

• Mt Staining by Indirect Immunofluorescence

• Ron Vale Mt Polymerization Buffer