Author
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Doss, Robert |
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Submitted to: Biotechniques
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 2/13/1996 Publication Date: N/A Citation: N/A Interpretive Summary: The differential display technique has been widely used to compare gene expression between two closely related populations of cells (for example, benign and malignant cells from the same organ or tissue). This publication describes modifications to the differential display technique that make it easier and safer to carry out. Among these modifications is the elimination of the requirement for radioactive compounds, and the use of a different enzyme for one of the steps. It is likely that these modificatons will be adopted by many laboratories when carrying out the differntial display procedure. Technical Abstract: A modification to the differential display technique is described. Silver staining rather than autoradiography is used to detect amplified cDNA bands, thus eliminating the need for radioactive materials. The Stoffel fragment of Taq polymerase is used for amplification, as it yields more products than the unaltered enzyme. Oligo d(T) is used for reverse transcription. Thus, only 2 reverse transcription reactions are needed. Either 8 or 24 reactions are required to obtain a representative population of cDNA if anchored primers are used. |
