Cloning in plasmids is usually accomplished by linearizing the plasmid with restriction enzymes, dephosphorylation, and gel-isolation. Ordinarily, this causes several problems that lead to decreased cloning efficiency. First, dephosphorylation decreases cloning efficiency approximately 50 fold. Ethidium bromide gel stain then forms adducts and intercalates the DNA, creating mutations and decreasing cloning efficiency. Ultraviolet light (both long and short wave) then degrades the DNA. Agarose inhibitors interfere with cloning efficiency. Finally, ‘shadow-band’ extraction allows residual uncut pDNA to survive the entire process, resulting in background (parental) plasmids.
NTC eliminates all of these issues, providing you with high-efficiency cloning vectors that have a 4-5 log reduction of background while retaining the cloning efficiency of plasmids that have not been gel isolated. This is perfect for high throughput screening, library production, and is a useful service for biotech suppliers who wish to provide their customers with highest quality cloning vectors, ready to use.
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