Absolute Quantitation of mRNA Species Using Northern Blotting, Nuclease Protection Assays and RT-PCR
Ambion's BrightStar™ Nonisotopic Detection System: Tips for Minimizing Background
Analysis of Differential Gene Expression
Here, we review three popular RNA detection/quantitation methods: Northern blot analysis, Ribonuclease Protection Assays (RPAs) and Reverse Transcription Polymerase Chain Reaction (RT-PCR). All of these can be used to confirm results from microarray analysis and differential display experiments.
Application of Prepared Liquid Phenols
Avoiding DNA Contamination in RT-PCR
BrightStar™ Psoralen-Biotin Labeled Probes
Cell Disruption: Getting the RNA Out
Choice of Ribonucleases for Ribonuclease Protection Assays
Determining RNA Probe Specific Activity and Yield
Gel Purification of Probes for Nuclease Protection Assays
Hang Up in the Wells - Preventing Samples from Remaining in the Wells of Your Gels
How to Maintain an RNase-free Lab
In Vitro Synthesis of RNA
Increasing Sensitivity in Northern Analysis with RNA Probes
Isolation of Total RNA from Difficult Tissues
Linearity of RNase Protection Assays
The experiment described in this Technical Bulletin demonstrates the use of Ambion's RPA III™ Kit to quantitate a target mRNA with a high degree of precision. It shows that this technique is capable of accurately detecting differences in target mRNA abundance that vary by as little as 20%.
Maximize DNA Probe Sensitivity
Membrane Transfer and Crosslinking for RNA
Methods for Nonisotopic Labeling of RNA by in Vitro Transcription
Methods to Remove DNA Contamination from RNA Samples
Nuclease and Protease Testing
Optimal Probe Concentrations for Ambion's Ribonuclease Protection Assay Kits
Optimizing In Situ Hybridization Protocols
Basic guidelines to simplify the choices and manipulations of various parameters for high quality ISH results.
Practical Tips for In Vitro Transcription
Practical Tips for Nonisotopic Detection
Practical Tips for Optimal Northern Analysis
Probe Specific Activity
Published siRNA Target Sequences
A handy list of siRNA target sequences that have been used in published gene silencing experiments. All siRNAs were used to silence genes in mammalian cells. Included in the list are links to GenBank, the siRNA sense and antisense strand sequences, and the appropriate template sequences for making each siRNA with the Silencer™ siRNA Construction Kit.
RNase and DEPC Treatment: Fact or Laboratory Myth?
Selected References on Microarray Analysis
Papers are categorized for easy reference. Categories include review articles, general interest, verification of results, instrumentation and software, aRNA amplification and useful web sites.
Selected References on RNA Structure/Function Analysis
Includes references (many are comprehensive review articles) on RNA structure analysis, commonly occuring RNA structures and how they are detected, and techniques to analyze protein-RNA interactions.
Simultaneous Detection of Multiple mRNA Targets with Ribonuclease Protection Assays
Simultaneous Extraction of RNA and DNA
siRNA Design Guidelines
A description of how to choose an siRNA target site, as well as how to design an appropriate hairpin sequence for use in expression vectors and cassettes.
The Accuracy of Competitive RT-PCR: Using the Right Exogenous Standard
The Basics of In Vitro Transcription
The Use of LiCl Precipitation for RNA Purification
Use of Internal and External Standards or Reference RNAs for Accurate Quantitation of RNA Levels
When Labeled Nucleotides Go Bad
Working with RNA