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United States Department of Agriculture

Agricultural Research Service

Title: Detection of Plant Genes Using a Rapid, Non-Organic Genomic DNA Isolation Method

Authors
item Lin, Jhy-Jhu - LIFE TECHNOLOGIES,INC.,MD
item Matthews, Benjamin
item Saunders, James
item Kuo, Jonathan - LIFE TECHNOLOGIES,INC.,MD
item Fleming, Ryan - LIFE TECHNOLOGIES,INC.,MD

Submitted to: Plant and Animal Genome Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: January 14, 1999
Publication Date: N/A

Technical Abstract: We have developed a simple procedure for the isolation of plant genomic DNA using FTA paper. Plant leaves were applied to FTA paper, and the genomic DNA was purified using a simple, non-organic reagent. The detection of 18S rRNA gene, and a gene [ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) gene] in chloroplast chromosome, was demonstrated using genomic DNA isolated from dicotyledonous plants such as poppy, tobacco, potato, Arabidopsis, soybean, and monocotyledonous plants, such as rice and corn, using FTA paper. Eight cultivars of soybean leaves were collected and stored on FTA paper for one month. Both the 18S RNA gene and the rbcL gene were detected in the genomic DNA isolated from these eight soybean cultivars. Furthermore, by increasing the number of cycles of DNA amplification, we are able to detect the GUS gene in transgenic tobacco and rice leaves using genomic DNA isolated by FTA paper. These results demonstrate that genomic DNA isolated using FTA paper can be used for the detection of plant genes (1) with either high or low copy numbers; and (2) located either in the nucleus or in an organelle. Further applications using genomic DNA purified by FTA paper will be discussed for the detection of single copy cDNA markers linked to a soybean cyst nematode resistance gene, for genetic analysis of forensic drug crops and for detection of AFLPs, PCR based DNA markers.

Last Modified: 4/18/2014
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