Abstract
DNA is the essential substrate for the polymerase chain reaction (PCR). Standard protocols include a DNA purification step, but this is laborious if a large number of DNA preparations have to be performed, although a variety of simple methods exist for the isolation of crude DNA for PCR. For microorganisms, PCR protocols exist that allow the amplification of sequences directly from the organism without DNA purification. The results introduced in this paper demonstrate that a direct PCR approach also works with fruitflies and blood flukes as examples for intact multicellular organisms.
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Selected References
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- Bier E., Vaessin H., Shepherd S., Lee K., McCall K., Barbel S., Ackerman L., Carretto R., Uemura T., Grell E. Searching for pattern and mutation in the Drosophila genome with a P-lacZ vector. Genes Dev. 1989 Sep;3(9):1273–1287. doi: 10.1101/gad.3.9.1273. [DOI] [PubMed] [Google Scholar]
- Clark A. G., Silveria S., Meyers W., Langley C. H. Nature screen: an efficient method for screening natural populations of Drosophila for targeted P-element insertions. Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):719–722. doi: 10.1073/pnas.91.2.719. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Collins J., Fleming T. Specific mRNA detection in single lineage-marked blastomeres from preimplantation embryos. Trends Genet. 1995 Jan;11(1):5–7. doi: 10.1016/s0168-9525(00)88975-1. [DOI] [PubMed] [Google Scholar]
- Finken M., Sobek A., Symmons P., Kunz W. Characterization of the complete protein disulfide isomerase gene of Schistosoma mansoni and identification of the tissues of its expression. Mol Biochem Parasitol. 1994 Mar;64(1):135–144. doi: 10.1016/0166-6851(94)90141-4. [DOI] [PubMed] [Google Scholar]
- Friedland L. R., Menon A. G., Reising S. F., Ruddy R. M., Hassett D. J. Development of a polymerase chain reaction assay to detect the presence of Streptococcus pneumoniae DNA. Diagn Microbiol Infect Dis. 1994 Dec;20(4):187–193. doi: 10.1016/0732-8893(94)90002-7. [DOI] [PubMed] [Google Scholar]
- Kaiser K. From gene to phenotype in Drosophila and other organisms. Bioessays. 1990 Jun;12(6):297–301. doi: 10.1002/bies.950120609. [DOI] [PubMed] [Google Scholar]
- O'Hare K., Rubin G. M. Structures of P transposable elements and their sites of insertion and excision in the Drosophila melanogaster genome. Cell. 1983 Aug;34(1):25–35. doi: 10.1016/0092-8674(83)90133-2. [DOI] [PubMed] [Google Scholar]
- Ramotar D. Rapid isolation of any known genes from whole cells of yeast by PCR. Mol Cell Biochem. 1995 Apr 26;145(2):185–187. doi: 10.1007/BF00935491. [DOI] [PubMed] [Google Scholar]
- Tirasophon W., Ponglikitmongkol M., Wilairat P., Boonsaeng V., Panyim S. A novel detection of a single Plasmodium falciparum in infected blood. Biochem Biophys Res Commun. 1991 Feb 28;175(1):179–184. doi: 10.1016/s0006-291x(05)81217-3. [DOI] [PubMed] [Google Scholar]
