Hello Fellow researchers.
I want to screen approx. 10^6 colonies from a mutant library that is supposed to show cellulolytic activity. I know that congo red and CMC in agar plate will form clear zone,but the wash and stain step will decrease the viability of cells. So i will not be able to pick the colonies showing positive for the cellulase test.
So are there any other alternatives to screen for celluloytic activity in petri dish and also pick the positive clones for further characterization, like use of fluorochrome dyes, where in the +ve or -ve colonies can be screened for enzyme activity and also can be picked from dish for further studies.
Kindly share me ideas and literatures if available
Hii...Goutham Genomics,
Screening of the mutant libraries for cellulolytic clones would be very laborious without high throughput screening system. However, various dyes are available now a days to screen the cellulolytic clones like Azure B ( AB ), Toluidene Blue O (TB) and Remazol Brilliant Blue R (RBBR) etc. that will help you to screen the cellulolytic clones on the plate itself. go through the attached paper..
Hello Yogesh thank you for your suggestion.
But my question is if i use any of the Dye based screening of clones, will i be able to pick up the +ve clones.? Like in congo red as there would be wash steps the possibility of getting viable cells is less. Similarly trypan blue was reported to be used in one paper where they picked the colonies showing to express endoglucanase activity forming halos. Do you feel this procedure of using trypan blue is viable for screening and also picking +ve clones for further characterization.?
Goutham Genomics,
Yes..Trypan blue also works well to screen the endoglucanase positive clones/ microbes from environmental samples and one can easily pick up the positive clones from the plates without any staining..
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