Dot blot assay technology: Low cost, non gel-based, high throughput technology for marker-assisted selection in cereals.
One of the marker-assisted selection methods being developed under GCP Project 18 is the dot blot based-genotyping assay. This method uses nylon membrane filters, and is therefore a gel-free and low-cost MAS application for cereals. To maximize the usefulness of this technique, we have increased the throughput of DNA templates that could be spotted on the nylon membrane, which would also contribute to cost reduction. The overall objective of the project is to develop, refine and validate allele-specific gel-free assays for biotic stress and quality traits in cereals that are low-cost assays and require low capital set-up and unit costs for NARES and small and medium enterprises. We have developed and validated this method using SNP markers based on the bacterial blight resistance gene Xa21 in rice and the Opaque 2 gene for quality protein maize.
Reveche, M.Y.; Mercado, E.; Skinner, D.; Carrillo, G.; Collard, B.; William, M.; Xu, Y.; Vera Cruz, C.M. Dot blot assay technology: Low cost, non gel-based, high throughput technology for marker-assisted selection in cereals. Presented at the 2006 GCP Annual Research Meeting, 12-16 September, Sao Paolo, Brazil. (2006) 1 pp.