Authors
Sook-Young Park, Myoung-Hwan Chi, Junhyun Jeon, and Yong-Hwan Lee
Summary
Rapid screening of mutant phenotypes is one of the integral parts in a large-scale study of gene functions. However, in filamentous fungi, strategy to address such a challenge has yet to be reported. Here we present our approach that allows rapid and systematic determination of multitude of phenotypes from insertional mutants of a filamentous fungus, Magnaporthe oryzae in a high throughput manner. To increase the speed and feasibility in screening phenotypes from a large collection of mutants, we developed high throughput phenotype screening (HPS) pipeline. Design of HPS pipeline was based on the fact that M. oryzae can undergo developmental changes away from its host, which allows visual and microscopic inspection of each developmental stage. This, together with the use of 24-well plate format, underlies our screening strategy. During HPS, mutant processing and evaluation of phenotypes were carried out in 24-well plate format. The use of 24-well plate greatly diminished the time and labor needed to label and handle every mutant because a single plate number automatically specifies twenty-four mutants within a plate and the twenty-four mutants go through HPS pipeline as a single unit.Further details
The protocol was published on Protocol Exchange in 2007. To see the entire protocol, click on the source link.Advertisement
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