Samarakoon, Tharangamala , Wang, Shiao , Alford, Mac .
Enhancing PCR amplification of DNA from recalcitrant plant species and herbarium specimens using a trehalose-based additive.
DNA amplification in plants is sometimes difficult due to the presence of phytochemicals that inhibit polymerase chain reaction (PCR). A reagent added to PCR, containing trehalose, bovine serum albumin and polysorbate-20 (TBT-PAR),has been used routinely for the detection of a virus in shrimp samples. The efficacy of using TBT-PAR to overcome PCR inhibitors was tested with DNA extracted from fresh, silica-dried, and herbarium leaf material that could not be PCR amplified using standard techniques. The addition of TBT-PAR resulted in amplification of all fresh or silica-dried samples and of a majority of herbarium samples, dating from 1989 to 2011. The addition of TBT-PAR during routine PCR is an effective method to improve DNA amplification of DNA extracted from herbarium specimens or plants that are known to have PCR inhibitors.
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1 - University Of Southern Mississippi, Department Of Biological Sciences, Alford Lab, 118 College Drive #5018, Hattiesburg, MS, 39406, USA
2 - University of Southern Mississippi, Department of Biological Sciences, 118 College Drive #5018, Hattiesburg, MS, 39406-0001, USA
Presentation Type: Oral Paper:Papers for Topics
Location: Union B/Hyatt
Date: Monday, July 9th, 2012
Time: 10:45 AM