Submitted to: Book Chapter
Publication Type: Book / Chapter
Publication Acceptance Date: August 1, 2005
Publication Date: January 1, 2006
Citation: Xin, Z., Chen, J. 2006. Extraction of Genomic DNA from Plant Tissue. In: Kieleczawa, J., Editor. DNA Sequencing II: Optimizing Preparation and Cleanup. 2nd Edition. Sudbury, MA: Jones and Bartlett Publishers. p. 47-59. Interpretive Summary: This book chapter reviewed the major methods used to extract genomic DNA from plant tissues. Emphasis is given on the principles of the methods, critical factors of success, and optimization procedures. It will help researchers to choose and optimize a method according to their individual research purposes.
Technical Abstract: Three general methods that are commonly used to extract genomic DNA from plant tissue were reviewed. The principles, critical factors, and optimization procedures were discussed to help researcher to choose and modify a method according to their research purpose. Emphasis is given to a simple 96-well plate based high throughput DNA extraction method that is applicable to many plant species. The method involves a simple incubation of plant tissue samples in a DNA extraction buffer followed by a neutralization step. With the addition of a modified PCR reaction buffer, the extracted DNA enabled the robust amplification of genomic fragments from samples of Arabidopsis, tobacco, sorghum, cotton, moss, and even pine needles. Several thousand DNA samples can be economically processed in a single day by one person without the use of robotics. This procedure will facilitate many technologies including high throughput genotyping, map-based cloning, and identification of T-DNA or transposon tagged mutants for known gene sequences.