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University of Washington School of Medicine, Department of Microbiology     

A. Tissue Culture (BL2) 
BL2 standard operating procedures .doc .pdf
TC supplies for Mullins lab .doc .pdf
Ten golden rules of tissue culture .doc .pdf
TC reagent recipes .doc .pdf
Effectene transfection protocol .doc .pdf
Liquid nitrogen freezer .doc .pdf
Thawing mammalian cells .doc .pdf
Freezing mammalian cells .doc .pdf
Passaging mammalian cells lines .doc .pdf
Mycoplasma PCR .doc .pdf
Isolation of PBMC using Ficoll gradients .doc .pdf
CD8 depletion by panning .doc .pdf
Magnetic cell sorting: Positive Selection .doc .pdf
Magnetic cell sorting: Untouched CD4 T+ cells .doc .pdf
Magnetic cell sorting: Untouched Monocytes .doc .pdf
X-Gal Staining of Transfected Cells .doc .pdf
B. Tissue Culture (BL3)
BL3 door sign .doc .pdf
BL3 standard operating procedures .doc .pdf
Processing HIV-1 infected blood .doc .pdf
Processing Sputum to obtain HIV-1 .doc .pdf
Biological cloning of HIV-1 .doc .pdf
Determining co-receptor usage (U87) .doc .pdf
Gradient Purification of AT2 Inactivated HIV-1 .doc .pdf
Isolation of Monocyte-Derived Macrophages from Buffy Coat by Plastic Adherence .doc .pdf
LAI Stock Preparation .doc .pdf
Virus concentration by PEG precipitation .doc .pdf
Virus growth curve .doc .pdf
Determining HIV-1 infectious titer .doc | .xls .pdf
HIV-1 LAI infection of T cell lines .doc .pdf
Tat stimulation of Jurkat T cells .doc .pdf
HIV-1 p24 antigen capture ELISA .doc .pdf
C. Flow Cytometry
Cell Quest User Guide .doc .pdf
Use of Rosen FACSCalibur .doc .pdf
FACSCalibur sign up sheet .doc .pdf
FACSCalibur user log sheet .doc .pdf
Monoclonal antibodies for FACS .xls .pdf
Detection of membrane antigens .doc .pdf
Detection of intracellular antigens (Caltag) .doc .pdf
Detection of intracellular antigens (BD) .doc .pdf
Detection of calcium flux .doc .pdf
Cell sorting by FACS .doc .pdf
D. Microarrays
Qiagen total RNA extraction .doc .pdf
Mullins Lab total RNA extraction .doc .pdf
Katze Lab total RNA extraction .doc .pdf
mRNA isolation by Oligotex (Qiagen) .doc .pdf
mRNA isolation by oligo(dT)-cellulose .doc .pdf
Eberwine T7 RNA amplification .doc .pdf
Preparation of linear acrylamide .doc .pdf
Bioanalyzer runs .doc .pdf
RNA-gel .doc .pdf
Radiolabeling DNA probe .doc .pdf
Hot asymmetric PCR .doc .pdf
Northern transfer .doc | .txt .pdf
Cy-Dye probe-synthesis .doc .pdf
Aminoallyl labeling .doc .pdf
Plasmid linearization .doc .pdf
In vitro transcription .doc .pdf
Quantitative PCR .doc .pdf
Reverse Transcription PCR .doc .pdf
Oligo hybridization for slide test .doc .pdf
First strand cDNA synthesis .doc .pdf
Processing of Q-bot membranes .doc .pdf
E. Other
Replication of cDNA clones (96-well) .doc .pdf
PCR of cDNA clones (96-well) .doc .pdf
Gel loading scheme .doc .pdf
Freezer Map Tool .doc .pdf
Purification of PCR products (96-well) .doc .pdf
Long PCR .doc .pdf
Whole Genome Subtype C Protocols .doc .pdf
TOPO T/A cloning of PCR products .doc .pdf
Real Time PCR .doc .pdf
Isolation of mammalian chromosomal DNA .doc .pdf
Big Dye terminator sequencing .doc .pdf
Sequence analysis .doc .pdf
Submission of 96-well plates for sequencing by the UW Biochem Sequencing Facility .doc .pdf
Protein gels .doc .pdf
14C mevalonic acid labeling .doc .pdf
14C acetic acid labeling .doc .pdf
 

 

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