zymolyase

In a spore culture, zymolyase (2 mg/ml) was used to digest ascum and liberate spores. We initially performed experiments to evaluate zymoliase sensitivity of asci for this specific strain and we assessed that 15 minutes incubation was the minimum time needed to damage the ascus, without touching the spore cell wall. Zymoliase was inactivated by heat (65°C for 2 minutes) and washed away carefully together with the remainings of the empty ascus, by resuspending twice the spore culture in 500 µl of distilled water centrifuging and discarding the supernatant; the spores were then resuspended to a concentration of 108 cells/ml. In order to exclude damage on the spore cell wall or effects on spore viability 100 µl of a culture of 1000 spores per ml was plated on YPD in triplicate, the spores showed 100% viability.

created over 14 years ago (21 January 2010)    last modified over 13 years ago (28 September 2011)   [ RDF ]   [ RelFinder ]