1Cereal Disease Laboratory, USDA, 2University of Minnesota/ Agroinnova, University of Torino, 3Cereal Disease Laboratory, University of Minnesota
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Pasquali, M., Kistler, C. Gibberella zeae Ascospore Production and Collection for Microarray Experiments.. J. Vis. Exp. (1), e115, doi:10.3791/115 (2006).
The protocol presented here is based on previous procedures used for perithecial production for Fusarium spp. (Klittich and Leslie, 1988; Trail and Common, 2000). Standardization of the procedure by which large quantities of ascospores (sufficient for microarray analysis) were generated was essential for the reproducibility of the experiment. It has been reported that environmental factors, age and substrate differences can change the biological character of ascospores (Beyer and Verreet, 2005). Therefore the use of video may highlight small details in the way spores are produced and collected that should facilitate reproducibility. In particular the video of the procedure should improve the level of standardization among laboratories and facilitate the comparison of whole genome transcription studies which require ascospore production. The amount of RNA necessary for experiment procedure is relatively high so a large number of Petri dish should be processed synchronously. Video is a particularly suitable tool when it is necessary to implement whole genome transcriptional studies on new biological material, setting a standard for future experiments.
The authors thank Karen Hilburn for excellent technical support. This project was supported by the National Research Initiative of the USDA Cooperative State Research, Education and Extension Service (Award #2004-35604-14327). Matias Pasquali is supported by Branco Weiss Fellowship. The US Wheat and Barley Scab Initiative is also acknowledged for the continuing support of research.
|Petri dish 9 cm Diameter||Tool|
|Tween 60 (20%)||Reagent|
|Carrot agar (20% w/v organic carrots and 1.5% w/v agar)||Reagent|
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