RAPID, UNIVERSAL METHOD TO ISOLATE PCR-READY DNA USING MAGNETIC BEADS

A magnetic bead-based system for DNA isolation utilizing monodisperse beads was tested with the aim of producing a general approach for PCR- ready DNA. This commercially available system was originally designed Sor isolating PCR-ready DNA from human whole blood. We rested dive,se organisms belonging to rite major groups bacteria, fungi, algae, vascu lar plants and vertebrates. Optimization of sample amounts and lysis c onditions was done using several types of tissue (fish epithelium, pla nt leaves, mammalian liver and muscle tissues, fungal fruit-bodies and mycelium). The standard lysis conditions used for blood could be appl ied with good results for most bacteria, algae and vertebrates, while plant leaves and fungal fruit-bodies had to be mechanically broken to obtain proper lysis. For vascular plants and some cyanobacteria, lysis by heating to 65 degrees C gave better DNA yields than standard lysis at room temperature. In all cases, DNA suitable for PCR was prepared in less than 30 min. The PCR products yielded 350 to 500 bases of DNA sequence (99% accurate) by direct manual or automated sequencing.