Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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Transfection Mediated by DEAE-Dextran: High-Efficiency Method

(Protocol summary only for purposes of this preview site)

Transfection mediated by DEAE-dextran differs from calcium phosphate coprecipitation in three important respects. First, it is used for transient expression of cloned genes and not for stable transformation of cells (Gluzman 1981). Second, it works very efficiently with lines of cells such as BSC-1, CV-1, and COS but is unsatisfactory with many other types of cells. Third, smaller amounts of DNA are used for transfection with DEAE-dextran than with calcium phosphate coprecipitation. Maximal transfection efficiency of 105 simian cells is achieved with 0.11.0 g of supercoiled plasmid DNA; larger amounts of DNA (>23 g) can be inhibitory. In contrast to transfection mediated by calcium phosphate, where high concentrations of DNA are required to promote the formation of a coprecipitate, carrier DNA is rarely used with the DEAE-dextran transfection method. For additional details on the procedure, see the information panel DEAE-Dextran Transfection.


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(Limited time special offer.) Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook
Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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