基因可隆的方法

 

• Serial Analysis of Gene Expression (SAGE) 
  SAGE is a powerful tool that allows the analysis of overall gene expression patterns with digital analysis.  Because SAGE does not require a preexisting clone, it can be used to identify and quantitate new gene as well as know genes. 
    
• cDNA/AFLP (CWB Bachem)
  The protocol which follows gives a step-by-step description of cDNA-AFLP, a method for visualisation of differential gene expression

・         What's Differential Display (GenHunter)
Introduction to differential display technique

・         Differential Display (Chun-Ming Liu)
The following procedures are described:

o        RNA extraction and qualification

o        DNase treatment of RNA sample

o        Reverse transcription

o        PCR amplification

o        Separation on acrylamide gels

o        DNA extraction from bands of interest

o        Re-amplification by PCR

o        Separation on agarose gels

o        Excision of amplified products
  

・         Differential Display (Breeden Lab)
Detailed protocol for differential display

・         Differential Display (Plant Molecular Biolgy Lab)
It's for plant RNA display. The protocol should be general.

・         Differential Display (PDF) (Clontech)
Provides detailed guide to differential display technique

・         Differential Display-Reverse Transcription-PCR (Gerard Lazo)

• Optimized Welsh DDRT-PCR Protocol (Gerard Lazo)
        
• Direct Characterization, Enrichment and Sequencing of Differential Displayed cDNA Sequences (TTO) 
  Present a rapid and cost-effective approach without recourse to cloning for direct characterization of differential cDNA molecules, up to the point of sequence identification. The experimental conditions have been optimized so that the protocol could be reliably performed with only three steps of minimal manipulation.
    

・         A simple method for screening cDNAs arising from the cloning of RNA differential display bands (TTO) 
This simple method allows to separate the obtained clones in subpopulations according to their sequence, thus reducing considerably the number of required reconfirmatory northern blot or reverse northern dot-blot reactions.

・         Rapid, nonradioactive differential display using Tth polymerase (TTO) 
A rapid method to produce RNA fingerprints that can be also used for differential display

・         Differential mRNA display using anchored oligo-dT and long sequence-specific primers as arbitrary primers (TTO)