Diabetes Genome Anatomy Project |
|
Resources > Protocols
Protocols
Project 1: Normal and Morbid Anatomy of Gene Expression in Mice
- Brown Fat Cell Isolation
- Genotyping of IRlox mice by PCR
- Genotyping of IR-heterozygous mice
- IRS-1 GENOTYPING BY PCR
- Genotyping of MCK-Cre mice by PCR
- Myocyte Isolation
- Measurement of Liver Triglyceride Content
Project 2: Identifying the underlying alterations in gene expression which result in type 2 diabetes
- RNA Isolation from Tissue Culture Cells w/ Trizol
- Procedure for RNA isolation from human muscle or fat
- cDNA & Biotinylated cRNA Synthesis Reactions for GeneChips
- Real-time quantitative RT-PCR ( Taqman )
Project 3: Anatomy of Gene Expression in Insulin Resistant States
- Isolation of Mouse or Rat Primary Fat Cells
- Glucose Uptake in Primary Fat Cells
- Assay for Free Fatty Acid Release from Isolated Fat Cells
- Assay for Conversion of U-14C-Glucose, 3H-Glycerol or 14C-pyruvate to Glyceride-Glycerol and Fatty Acids in Isolated Fat Cells
- RNA Isolation From Primary Fat Cells using Trizol from Invitrogen
Project 4: Information of levels, modification states and subcellular distribution of proteins in cells and tissues
- Isolation and Analysis of sub-cellular Fractions from 3T3L1 Adipocytes
- Isolation and Analysis of Sub-cellular Fractions from Small and Large Adipocytes from Wildtype and Firko Mice
Project 5B: Genetics of Adipose-Regulated Genes in Insulin-Resistance and Type 2 Diabetes
Project 7: Interfacing Small Molecules with the Diabetes Genome Anatomy Project
Project 8: Signal-dependent Interactions in Insulin Action
- Project_8_Mann.doc contains:
- Culture of brown adipocytes (BAT) in SILAC Medium and Insulin induction
- Immunoprecipitation
- In Gel digestion
- Sample preparation after "in gel" digestion
- HPLC system and mass spectrometer
- Relevance networks protocol - Introduction to Relevance Networks by Atul Butte, M.D., Ph.D.
- Existing methods protocol - Introduction to existing methods in clustering microarray data by Atul Butte, M.D., Ph.D.
- Amia Tutorial 2003 - Introduction to Bioinformatics tutorial slides presented at American - Medical Informatics Association 2003 meeting by Atul Butte, M.D., Ph.D.