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blank and substrate control give OD reads


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#1 intandiana

intandiana

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Posted 15 January 2014 - 12:03 AM

Hi there..smile.png

I'm working on screening of xylanase variants to look for the better variant than the wild type in hydrolysis kenaf. the reaction is done in 96 deep well plate by adding substrate (kenaf) + buffer (50mM sodium acetate, pH 5.0) + enzyme solution and incubate for 4 hours in waterbath. after 4hrs incubation, the reaction is centrifuged to collect only reaction solution (without the substrate) then assay the reaction with DNS and incubate at 100oC, 5min. each experiment is controlled by blank (buffer only), substrate (buffer + substrate) and wild type xylanase.

the problem is the blank and substrate control give higher OD reads than the wild type and variants. I added antimicrobial (tetracyline-HCL) into the reaction mixture but not work. wacko.png

any suggesstion to help me??blink.png






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