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Purification of Thermostable DNA Polymerases

Thermostable polymerases are of course key enablers of molecular biology, due to their usefulness in polymerase chain reaction (PCR).  Today I am going to show you the process by which these important enzymes are isolated.  You will learn how to make your own Taq polymerase using the open source expression vector pOpenTaq, as well as understand the purity grades of the commercial polymerases offered by Gene And Cell Technologies.  The principles outlined in this blog post should apply to any thermostable nucleic acid polymerase.   Taq polymerase can be expressed form pOpenTaq, an open-source taq polymerase expression plasmid.  pOpenTaq should..  - Read More

DNA Purification from Agarose Gels

  By John Schloendorn   Introduction Over the past couple of years, I had the good fortune to learn a whole lot about DNA, while assembling Gene And Cell Technologies’ molecular armamentarium from my own genome and a variety of other sources.  Today I’d like to highlight one particular aspect of the process, the extraction of DNA fragments from agarose gels for subsequent cloning.   Cloning experiments of course require excellent quality DNA fragments, absent both DNA and non-DNA contaminants.  The kinds of molecules that need to go include damaged plasmids, uncut plasmids, PCR template carryover, false PCR products, primer..  - Read More