Deoxyribonucleic Acid Preparation in Polymerase Chain Reaction Genotyping of Transgenic Mice
Abstract:Background and Purpose: In an attempt to find a rapid and reproducible method for routine polymerase chain reaction genotyping of transgenic mice, two novel approaches were developed.
Methods: One approach allowed reproducible amplification from crude lysates of tail snips, using a thermally activated polymerase. In a second approach, for situations in which non-invasive techniques are necessary, oral swab specimens were amplified after DNA extraction by use of an isolation kit. Samples from 10 transgenic factor VIII knockout mice were genotyped after processing by use of these and other methods.
Results: False-negative results were not obtained by use of the two novel approaches. Despite their relative simplicity, both approaches yielded results comparable to those obtained by use of procedures known to be reliable, such as organic extraction and a DNA extraction kit.
Conclusion: Both approaches are useful for PCR-amplification of DNA from mammalian sources.
Document Type: Research Article
Publication date: 2000-06-01
Comparative Medicine (CM), an international journal of comparative and experimental medicine, is the leading English-language publication in the field and is ranked by the Science Citation Index in the upper third of all scientific journals. The mission of CM is to disseminate high-quality, peer-reviewed information that expands biomedical knowledge and promotes human and animal health through the study of laboratory animal disease, animal models of disease, and basic biologic mechanisms related to disease in people and animals.
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