Quantitative digital image analysis of chromogenic assays for high throughput screening of alpha-amylase mutant libraries.
An image analysis-based method for high throughput screening of an alpha-amylase mutant library using chromogenic assays was developed. Assays were performed in microplates and high resolution images of the assay plates were read using the Virtual Microplate Reader (VMR) script to quantify the concentration of the chromogen. This method is fast and sensitive in quantifying 0.025-0.3 mg starch/ml as well as 0.05-0.75 mg glucose/ml. It was also an effective screening method for improved alpha-amylase activity with a coefficient of variance of 18%.
Shankar
Manoharan
M
Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, Madurai, 625021, India.
Priyadharshini
Ramachandran
R
Gunasekaran
Paramasamy
P
D002863:Chromogenic Compounds / Q000378:metabolism*; D019020:Directed Molecular Evolution; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D000516:alpha-Amylases / Q000235:genetics* / Q000378:metabolism*
D002863:Chromogenic Compounds; D000516:alpha-Amylases