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Welcome to the Bridges Lab protocol site. If you would like to know more about our group, please visit the [http://bridgeslab.sph.umich.edu Bridges Lab Website]. All of our experimental protocols are available through this site and are free to use/copy/modify by anyone without any restrictions. You can locate protocols by looking at this index, searching by category (see below) or searching by keyword. If you notice an error, would like to discuss these methods or would like to be able to edit these pages contact us via email.
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==Protocol Categories==
===Cell and Tissue Culture===
*[[Splitting Cells]]
*[[Generating DMSO Stocks for Cell Culture]]
*[[Inositol Labeling and Lipid Extraction]]
===Cloning and Molecular Biology===
*[[Transformation of Bacteria]]
*[[Mutagenesis]]
*[[Preparing an Agarose Gel]]
*[[Cesium Chloride Preparation of DNA]]
*[[Designing and Generating CRISPR-Cas Mutants]]
===Cell Biology===
*[[Immunofluoresence]]
**[[Colocalization]]
*[[FM4-64 Labeling of Yeast Cells]]
===Protein Analysis===
*[[Preparing a SDS-PAGE Gel]]
*[[Western Blotting]]
*[[Surface Plasmon Resonance - Protein Lipid Interactions]]
====Protein Quantification====
*[[Bradford Assay]]
*[[Quantification by Absorbance at 280nm]]
*[[Using ImageJ to Quantify Bands]]
====Protein Purification====
*[[French Press]]
*[[Purification of GST Fusion Proteins]]
*[[Glycogen Synthase Assay]]
*[[Triglyceride Assay from Cells and Tissues]]
*[[Triglyceride Assay from Drosophila (German Method)]]
===Transcriptional Analysis===
*[[Real Time PCR From Cell Culture]]
*[[Harvesting RNA from Cells grown in monolayer]]
*[[Using Bioconductor To Analyse Microarray Data]]
===Mouse /Fly Protocols===*[[Maternal Particulate Exposure Breeding]] ===Fly Stocks===*[[Maintenance of 18C Fly Stocks]]*[[Fly Food Protocol]]
===Genotyping===
*[[Ear Tagging and Tail Clipping]]
*[[PCR Analysis of Tail DNA]]
*[[Colony PCR]]
*[[BDNF PCR]]
*[[TaqMan SNP Assay]]
===Metabolic Measurements===
*[[Monitoring Food Intake]]
===Tissue Preparation===
*[[Harvesting Mouse Tissue]]
*[[Preparation of Protein Lysates from Mouse Tissues]]
*[[Preparation of RNA Samples from Mouse Tissues]]
*[[Paraffin Embedding of Tissue Samples]]
*[[H&E Staining of Mouse Tissue]]
**[[Adipocyte Cell Counting with ImageJ]]
*[[Perfusion of Mouse]]
===Media and Buffers=======Yeast Media====
*[[YPD Media and Agar]]
*[[Yeast Selective Media and Agar]]
====Buffers=========Lysis Buffers=====
*[[KRBH Buffer]]
*[[RIPA Buffer]]
*[[CHAPS Lysis Buffer]]
=====SDS-PAGE Buffers=====
*[[SDS-PAGE Separating Gel Buffer]]
*[[SDS-PAGE Stacking Gel Buffer]]
*[[TORC1 Kinase Buffer]]
*[[Acrylamide Solution]]
=====Other Buffers=====
*[[KRBH Buffer]]
*[[MTORC1 Kinase Assay]]
