Construction of a synthetic vector for preparation of a 100 base pair DNA ladder

Abstract

DNA size markers are widely used to estimate the size of DNA samples on agarose or polyacrylamide gel<br />electrophoresis (PAGE). DNA markers can be prepared by mixing PCR products with definite sizes.<br />Alternatively, they are prepared by restriction enzyme digestion of the genomic DNA of bacteriophages or<br />natural and synthetic DNA plasmids. The present study describes engineering of a synthetic plasmid<br />which produces a 100 bp DNA ladder, a popular DNA size marker, upon digestion with a single restriction<br />enzyme. Our strategy consisted on sequential cloning of ten PCR products of 100 to 1000 bp in plasmid<br />pTZ57R, using the BamHI and BglII restriction enzymes and releasing the fragments from the recombinant<br />plasmid by enzyme EcoRV. This strategy could be applied to construct various complex synthetic vectors<br />to produce different DNA ladders.