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The use of TaqMaster PCR Enhancer

Guy Adami, Department of Oral Medicine, University of Illinois in Chicago, USA
Vincent Prezioso, Eppendorf Scientific, Inc., Westbury, NY, USA

Introduction

Transgenic mice carry a fragment of foreign DNA that has been incorporated into their genome. The most common method of introducing recombinant DNA into the mouse genome is by microinjection of the DNA into the pronuclei of one-cell stage mouse embryos.

A fraction of these embryos develop into newborn mice which have the micro-injected DNA stably integrated in their genome. A second strategy for generating transgenic mice involves genetic manipulation of ES cells.

After electroporation with a carefully designed construct, the stem cells are screened for the integration of new DNA through homologous recombination.

ES cells that are positive for a homologous recombination event are then transplanted into blastocyst stage embryos in order to generate chimeric mice. In most cases, gene targeting is used to introduce mutations into specific genes in the mouse genome.

The transgenic DNA is typically transcribed and translated in a tissue-specific and stage-specific pattern that is determined by regulatory sequences present in the transgenic DNA. Consequently, transgenic mice represent an experimental system in which it is possible to study the consequences and/or pattern of expression (or lack thereof) of the new genetic information. Incorporated transgenes are often detected by genomic DNA isolation from a tissue sample, followed by PCR* amplification of a region of the inserted
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