Bottone Lab Protocols

 

Dr. Frank G. Bottone, Jr.

The University of North Carolina at Chapel Hill  

 

 

 

Bottone Lab Protocols

 

Molecular Biology Protocols for Fellow Lab Rats I have developed most if not all of these protocols while at NIEHS, UNC, or Duke. They all work well for me, but be sure to get additional information if you are not familiar with the Intricacies of them.

 

Tissue Culture

  1. Cell lines and reagents I have used

  2. FREEZING CELLS

  3. TYPE II CELL PREP FROM RAT LUNGS

  4. MONOCYTE ISOLATION

  5. PERIPHERAL BLOOD CELL ISOLATION

  6. ROSETTE

  7. COLORECTAL CELL LINES

  8. ISOLATION OF PIG CELLS

 

Enzymes Stuff

  1. COX-2 Semiprotein Preparation

  2. Oxygraph Setup

 

DNA Techniques

  1. EMSA

  2. PCR VENT TAQ

  3. PCR PLATNIUM TAQ

  4. SYBR GREEN PCR

  5. RT SYBR GREEN

  6. Commonly used dual labelled probes.html

  7. DIG RNA

  8. ELECTROPHORESES

  9. LABELING

  10. OLIGO PREPERATION

  11. SEQUENCING SETUP

  12. SEQUENCING GELS

  13. TOPO Cloning

  14. Transformation

 

RNA and Reverse Transcription

  1. SUPER SCRIPT II RT

  2. SUPER SCRIPT RT

  3. QIAGEN OMNISCRIPT RT

  4. NORTHERN BLOTTING

  5. RNA ISOLATION THE OLD FASHION WAY

  6. RIBONUCLEASE PROTECTION ASSAY

 

Proteins

  1. VEGF ELISA Using Kit

  2. VEGF ELISA Quantikine.pdf file

  3. PAGE

  4. BCA MICROPLATES

  5. Whole Cell Lysates from Monolayer Cultures

  6. Cell membrane preparation

  7. PAGE NOVEX

  8. PROTEIN ISOLATION FROM TISSUE

  9. PROTEIN ISOLATION FROM MONOLAYER

  10. PROTEIN ISOLATION FROM TRIZOL

  11. SDS PAGE

  12. Nuclear Extracts

  13. WESTERN BLOTTING

 

In situ

  1. tunnel staining for Apoptosis

  2. H and E staining

 

Other Stuff for My Laboratory

  1. SUPPLIES

  2. LIST OF REAGENTS

  3. restriction table.html

  4. HPLC