Plant & Animal Genomes XI Conference Plant & Animal Genomes XI ConferenceJanuary 11-15, 2003
Town & Country Convention Center
San Diego, CA
Poster: High-throughput Methods
The Wizard Magnetic 96 DNA Plant System was designed for DNA purification from commercially important agricultural crops such as corn and tomato leaf as well as canola and sunflower seeds, with sample sizes of one 8 mm diameter leaf punch or 1-5 seeds. About 50 - 100 ng DNA is obtained. Many other plant species have been tested also. Plant material is ground in the presence of lysis buffer in a 96 deepwell plate. After centrifugation to pellet debris, the lysate is transferred by the robot to another plate containing Magnesil particles which bind DNA. After washing steps the particles are briefly dried and the DNA eluted in TE buffer or water. The protocol was initially set up on a Biomek 2000 liquid handling system (Beckman Coulter Instruments) with a gripper arm to transfer plates on the deck. We have adapted this to increased DNA recovery on a Biomek FX, which allows 96 isolations to be done simultaneously with larger sample sizes. In our experience, from 500 ng to 2 ug DNA can be obtained depending on sample species. An alternate method to obtain a fixed yield from each sample type eliminates the need to measure DNA for downstream testing. The DNA obtained can be used in PCR, RAPD analysis and ISSR testing.