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[13] Preparation of DNA from blood for polymerase chain reaction in microtiter dish

Elsevier Science & Technology
DOI: 10.1016/0076-6879(92)16015-c
  • Section I. Isolation
  • Synthesis
  • And Detection Of Dna And Rna
  • Biology
  • Design


Publisher Summary The preparation of DNA is often a rate-limiting step in obtaining a PCR product from a biological source. A strategy was devised to minimize the number of manipulations and to obviate the need for centrifugation. Anti-histone antibodies are adsorbed to a tube or well of a microtiter dish. This technique has been designated immuno-PCR. Although one believes this technique to have general applications, the example described in the chapter is the preparation of plasmodium falciparum DNA from malaria-infected human blood. The chapter discusses used anti-histone antibodies to capture the chromatin but other agents could be used to capture DNA. These include antibodies to control elements that bind DNA for prokaryotes, antibodies to DNA itself, and antibodies to the target micro-organisms itself. Ligands other than antibodies could be used, e.g., nonspecific DNA-binding proteins. Given the experience with immuno-PCR, a critical factor to investigate in these systems would be the buffer composition.

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