cDNA Library Construction sans Restriction Enzymes

Complementary DNA (cDNA) libraries are standard tools for gene expression studies. In theory, the library contains full-length DNA copies of every mRNA in the starting sample, in abundances representative of the original sample. In reality, however, the libraries are often missing rare clones and contain partial gene fragments, complicating subsequent analyses. Further, transferring specific cDNA clones from one vector to another--to express the protein in mammalian cells instead of in bacteri

Jeffrey Perkel
Mar 23, 2003

Complementary DNA (cDNA) libraries are standard tools for gene expression studies. In theory, the library contains full-length DNA copies of every mRNA in the starting sample, in abundances representative of the original sample. In reality, however, the libraries are often missing rare clones and contain partial gene fragments, complicating subsequent analyses. Further, transferring specific cDNA clones from one vector to another--to express the protein in mammalian cells instead of in bacteria, for example--can be hampered by the presence of inopportune restriction enzyme cut sites within the cDNA sequence. A new product from Carlsbad, Calif.-based Invitrogen addresses both of these problems.

The CloneMiner™ cDNA library construction kit employs two Invitrogen technologies: SuperScript™ II reverse transcriptase, and Gateway recombination-based cloning. Gateway is based on lambda phage recombinase; the enzyme, which Invitrogen calls Clonase™, swaps DNA fragments flanked by att sites, making it possible to move fragments between Gateway- compatible plasmids without restriction...

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