Fission Yeast

Biochemistry

Molecular Biology

Tissue Culture/Cell Biology

Bioinformatics

Others

• Nurse lab manual
• Expressing protein from pRep transformed yeast
• Preparation of cell lysate for SDS-PAGE (S. Wu)
• Isolation of diploids (general) (R. Geyer)
• Isolation of diploids (specific) (R. Geyer)
• Genomic C-terminal tagging by PCR (C. Zhou)
• Fractionation of cytoplasmic and nuclear proteins (V. Seibert)
• FACS (V. Seibert)
• Immunofluorescence by formaldehyde fixation (E. Rhee)
• Antibiotic sensitivity (E. Rhee)
• Preparation of genomic DNA for PCR
• Determining protein stability in vivo by cycloheximide chase (S. Wee)
• Freezing S. pombe cells (S. Wee)
• Calcofluor staining (F. Arslan)
• Preparation of S. pombe lysate for LC-MS/MS analysis (S. Wu)
• Large scale S. pombe cell lysate preparation for biochemistry - Coffee grinder method (E. Amoah-Buahin)
• Making haploids from Bioneer h+/h+ diploids (S. Wu)
• Polysome fractionation and mRNA extraction (P. McQuary)

• Bacterial Expression in BL21
• Flex column assembly and use
• Concentrating protein
• Affinity Purification of antibodies (V. Seibert)
• Purification of monoclonal antibody from hybridoma supernatant (C. Zhou)
• Gradient minigels
• Large gradient gels (S. Wee)
• Vorum silver staining protocol (compatible with MALDI MS)
• Fractionation of cytoplasmic and nuclear S. pombe proteins
• In-gel digestion of proteins for MALDI MS
• Multidimensional 2D gel analysis of S. pombe lysate
• Affinity purification of protein complexes with IgG/proA (Version 1 - R. Geyer)
• Affinity purification of protein complexes with IgG/proA (Version 2 - Optimization by Z. Sha)
• TEV cleavage and TCA precipitation
• In vitro ubiquitination assay with IPed cullin complexes (M. Schmidt)
• Autoinduction of fusion proteins in BL21 (E. Rico-Bautista)
• Tris-Tricine gels for small proteins (E. Rico-Bautista)
• Preparation of S. pombe lysate for LC-MS/MS analysis (S. Wu)
• Affinity purification of GFP fusion proteins from S. pombe or mammalian cells (J. Scheliga)
• Polysome fractionation and mRNA extraction (P. McQuary)
• FLAG purifications from transfected mammalian cells (J. Scheliga)
• Polysome profiling with mammalina cells (DAW)

• Competent bacteria
• Miniprep
• RNA preparation and RT-PCR (S. Wee)
• Northern Blotting
• Yeast 2H screening (A. Piau)

• Purification of monoclonal antibody from hybridoma supernatant
• Production of baculoviruses
• Expression analysis of recombinant proteins
• Production of adenoviruses (H. Schulz)
• Amplification of adenoviruses (C.C. Yang)
• Immunofluorescence staining of LNCaP prostate cancer cells (E. Rico-Bautista)
• FACS (D.A. Wolf)
• High efficiency transfection of LNCaP and PC-3 cells (C.C. Yang)

• Assembling gene lists from InterPro protein domains (also includes gene name conversion with Panther) - (C.-C. Yang)

• Sporulation of budding yeast (S. Wee)
• Transformation of budding yeast (S. Wee)