|Lin, Jhy-Jhu - LIFE TECHNOLOGIES, INC|
|Fleming, Ryan - LIFE TECHNOLOGIES, INC|
|Kuo, Jonathan - LIFE TECHNOLOGIES, INC|
Submitted to: Plant and Animal Genome Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: November 9, 1998
Publication Date: N/A
Technical Abstract: We 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 dicotyledous plants such as tobacco, potato, Arabidopsis, soybean and monocotyledous plants such as rice and corn using FTA paper. Soybean leaves from eight cultivars 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 detected 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 and for detection of AFLP , a PCR based technique for identifying DNA markers.