|Lin, Jhy-Jhu - LIFE TECH., ROCKVILLE, MD|
|Fleming, Ryan - LIFE TECH., ROCKVILLE, MD|
|Kuo, Jonathan - LIFE TECH., ROCKVILLE, MD|
Submitted to: Plant Genome Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: January 14, 2001
Publication Date: N/A
Technical Abstract: A new procedure for the rapid extraction and storage of DNA from plant tissue has been developed called FTA DNA isolation. In a previous report, we showed that this technology was capable of rapidly extracting DNA from plant leaves in a form that was suitable for PCR screening. Large numbers of plant samples could be processed with minimal effort, and the FTA cards containing the plant DNA can be stored for several years at room temperature without sample degradation. Due to the presence of high levels of endogenous secondary natural products in some plants, a small number of replicate samples did not show positive PCR amplification during screening protocols. We revised sample processing protocols to produce improved results in plants with high levels of endogenous inhibitors. The improved methodology involves changes in tissue disruption producers and modifications of the FTA DNA extracting buffers for PCR amplification. Using these revisions, FTA DNA isolation procedures can be used in wider number of plants, regardless of their endogenous biochemical makeup.