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Improving DNA amplification from problematic plants
Date:1/3/2013

The polymerase chain reaction (PCR) is a common technique used to amplify, or copy, pieces of DNA. Amplified DNA is then used in genetic analyses for everything from medicine to forensics. In plant research, PCR is a vital step in detecting and sequencing genes, and its applications are endless. However, compounds found in plants often inhibit PCR. Researchers at the University of Southern Mississippi discovered that the use of an additive allows PCR to successfully amplify DNA from once problematic plants.

PCR is widely used in plant sciences but is not 100 percent reliable. Many plant researchers encounter roadblocks when implementing PCR. For example, many plant species contain phenolic compounds that deter herbivores. These compounds are often extracted along with plant DNA and can stop PCR from working.

Graduate student Tharangamala Samarakoon and colleagues have found a technique to overcome many of these inhibitory plant compounds. They added a reagent to the PCR mixture that contains three ingredients: trehalose, bovine serum albumin, and polysorbate-20 (all three abbreviated TBT-PAR). "Unlike several other studies, TBT-PAR works at the PCR stage instead of at the DNA extraction stage, so it has promise for pigeon-holed and half-forgotten extractions that previously failed to be amplified using PCR," says Samarakoon. The authors published their research in the January issue of Applications in Plant Sciences.

Samarakoon tested the TBT-PAR reagent on DNA extracted from tropical and temperate species across four plant families, including Achariaceae, Asteraceae, Lacistemataceae, and Samydaceae. PCR with TBT-PAR successfully amplified DNA for all species, whereas standard DNA extraction and PCR techniques consistently failed.

TBT-PAR enhanced PCR for DNA extracted from fresh, silica-dried, and herbarium plant material. "Si
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Contact: Beth Parada
bparada@botany.org
American Journal of Botany
Source:Eurekalert

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