Protocols and Applications Guide

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Download individual chapters using the links in each chapter or download the entire Guide using these links (14MB): Letter or A4.

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Each chapter includes:

Step-by-step protocols
Illustrations and example data
Extensive background information
Animated movies of key techniques and cellular events
Links to citations and other useful laboratory tools

GUIDE CONTENTS

PCR Amplification
PCR Applications
- Introductory animation about PCR
- Optimization tricks
- RT-PCR tips
- Polymerase and transcriptase review
- PCR & RT-PCR protocols
- Troubleshooting
RNA Interference
RNA Interference
- Learn how to design and optimize siRNA sequences
- Use reporter genes for siRNA optimization
- Investigate RNAi delivery strategies
- View a Nature animation about the RNAi mechanism
Apoptosis
Apoptosis
- Gather more data from a single sample with multiplexed assays
- Use two or more methods to confirm apoptosis
- Detect caspase activity with anti-PARP pAb
- Learn about the molecular mechanisms of apoptosis
Cell Viability
Cell Viability
- Compare assay methods based on your needs
- Read about factors that effect viability assay outcome
- Learn how to multiplex cell viability with cytotoxicity or reporter assays
- Choose an assay with the Cell Viability Assistant
Protein Expression
Protein Expression
- Optimize your protein yield
- Learn about the gold standard: TNT® Systems
- Use the product selector tool to choose your system
- Explore eukaryotic, mammalian and bacterial systems
- Read about non- radioactive labeling systems
Expression Analysis
Expression Analysis
- Single gene or gene array systems
- Tips for creating a ribonuclease-free environment
- Recipes for common solutions
- Total or polyA RNA isolation systems
Cell Signaling
Cell Signaling
- Animations of MAPK and PI-3K pathways
- High or low ATP concentration kinase asssays
- Fluorescent, luminescent, radioactive and colorimetric kinase assays
- Phosphorylation-specific antibodies for MAPK pathways
Bioluminescent Reporters
Bioluminescent Reporters
- pGL4: The latest generation reporter vectors
- Dual-luciferase system: for better control
- Live-cell substrates

DNA Purification
DNA Purification
- Tips to maximize yield and quality
- Plasmid and genomic DNA purification
- Transfection-quality miniprep DNA
- High-, low- and medium-throughput
- DNA from blood
- Automated purification at your lab bench
Cell Imaging
Cell Imaging
- HaloTag® Technology: One construct, multiple fluors
- Live-cell imaging protocols
- Rapid labeling and imaging
- FACS®
Protein Purification and Analysis
Protein Purification and Analysis
- Affinity tags for purification
- Analyzing DNA:Protein interaction
- HaloCHIP™ System
- Pulldown assays for protein interactions
- Recipes for commonly used solutions
Transfection
Transfection
- Methods for achieving transfection
- Optimizing transfection efficiency
- Sample transfection protocol
- Recipes for commonly used solutions
Cloning
Cloning
- Learn about T-Vector cloning
- Use Flexi® Vectors for easy sequence transfer from one vector to another
- Get technical information on restriction enzymes
- Find out what affects PCR cloning
- Recipes for solutions used in cloning
DNA-Based Human Identification
DNA-Based Human Identification
- Learn about how repetitive DNA is used for human identification applications such as forensic casework and relationship testing
- Explore the steps required to generate a DNA profile
- Gain information about automated key steps in the DNA-typing process
- Discover links to Promega protocols, Profiles in DNA articles, validation guides and other resources
Buffers for Biochemical Reactions
Buffers for Biochemical Reactions
- Explore how a buffer system works
- Understand what makes a "good" buffer
- Learn how to prepare buffers
- Find recipes for common buffers
Epigenetics
Epigenetics
- Explore epigenetic mechanisms of gene regulation
- Discover how DNA methylation and histone modifications affect chromatin structure
- Decipher the histone code
- Learn about techniques to research epigenetics