European Journal of Forest Pathology (1996) 26, 81-88
A. Pappinen, R. Kasanen, T. Möykkynen and K. von Weissenberg (1996)
Fast methods for DNA extraction for PCR analyses from Heterobasidion annosum mycelium and Endocronartium pini aeciospores
European Journal of Forest Pathology 26 (2), 81-88
Abstract: The extraction of DNA from mycelium of Heterobasidion annosum or aeciospores of Endocronartium pini could be done rapidly by homogenizing the fungal material in PCR-compatible buffer in Eppendorf tubes with a disposable pestle mounted on an electric drill. The DNA yield with this method was relatively low but sufficient for PCR analysis with ITS-specific primers. For large series of fungal samples this protocol provides a quick and cheap method of determining the intersterility groups of H. annosum or for specific primer studies on E. pini populations. The time needed for DNA isolation from E. pini aeciospores and H. annosum mycelium could be reduced by nearly 85% compared to the mortar-and-pestle protocol.
By using a disposable pestle mounted on an electric drill and phenol-chloroform as extraction solution, the yield of DNA was higher than with the mortar-and-pestle method with phenol-chloroform as extraction solution. The quality of DNA was sufficient for both specific-primer and RAPD analyses, but the more complicated extraction method with phenol was needed.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Link to article at publishers website
Database assignments for author(s): Timo Möykkynen
Research topic(s) for pests/diseases/weeds:
molecular biology - genes
Pest and/or beneficial records:
Beneficial | Pest/Disease/Weed | Crop/Product | Country | Quarant.
|
---|---|---|---|---|
Heterobasidion annosum | ||||
Cronartium flaccidum |