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Fission Yeast
Biochemistry
Molecular Biology
Tissue Culture/Cell Biology
Bioinformatics
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Fission Yeast
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)
Biochemistry
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)
Large scale S. pombe cell lysate preparation for biochemistry - Coffee grinder method (E. Amoah-Buahin)
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)
Molecular Biology
Competent bacteria
Miniprep
RNA preparation and RT-PCR (S. Wee)
Northern Blotting
Yeast 2H screening (A. Piau)
Tissue Culture/Cell Biology
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)
Bioinformatics
Assembling gene lists from InterPro protein domains (also includes gene name conversion with Panther) - (C.-C. Yang)
Others
Sporulation of budding yeast (S. Wee)
Transformation of budding yeast (S. Wee)