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ARS Home » Plains Area » Lubbock, Texas » Cropping Systems Research Laboratory » Plant Stress and Germplasm Development Research » Research » Publications at this Location » Publication #131999
Title: A SIMPLE VERSATILE HIGH THROUPHPUT DNA EXTRACTION METHOD SUITABLE FOR PCR

Author
item Xin, Zhanguo
item Velten, Jeffrey
item Oliver, Melvin
item Burke, John

Submitted to: American Society of Plant Biologists Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 2/6/2003
Publication Date: 4/20/2003
Citation: Xin, Z., Velten, J.P., Oliver, M.J., Burke, J.J. 2003. A simple versatile high throughput DNA extraction method suitable for PCR. American Society of Plant Biologists Annual Meeting. 34(4):820-826.

Interpretive Summary:

Technical Abstract: PCR has become the most popular technique in functional genomics. Both forward and reverse genetic projects routinely require PCR amplification of thousands of samples. Processing the samples for DNA suitable for PCR is usually the limiting step. We have developed a simple high throughput DNA extraction method that is applicable to many plant species. The method involves a simple incubation of plant sample in a 96-well PCR plate in a DNA extraction solution followed by neutralization. With a modified PCR buffer, the DNA extracted with our method allowed robust amplification for pine needles, cotton leaves, mosses, and Arabidopsis. Several thousands DNA samples can be processed in a day by one person. This method will aid the high throughput genotyping and identification of T-DNA or transposon tagged mutant for a known gene. The critical factors for the successful application of this method will be discussed.