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Preparations of the fungal CW and EPS matrix were incubated overnight in sterile Milli-Q water at 65 °C prior to enzymatic digestion. Substrate and enzyme concentrations, buffer compositions, digestion temperature and time are described in the figure captions. Digestion was stopped by denaturing the enzymes at 95 °C for 10 min and the digestion products were stored at -20 °C prior to use. \ No newline at end of file diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/TLC.md b/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/TLC.md new file mode 100644 index 0000000..a700062 --- /dev/null +++ b/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/TLC.md @@ -0,0 +1,2 @@ +## Thin layer chromatography (TLC) +An aliquot of each sample was subjected to TLC using a silica gel 60 F254 aluminum TLC plate (Merck Millipore, Burlington, USA), using a running buffer containing ethyl acetate/acetic acid/methanol/formic acid/water at a ratio of 8:4:1:1:1 (v/v). D-glucose, laminaribiose β-1-3-(Glc)2, laminaritriose β-1-3-(Glc)3, gentiobiose β-1-6-(Glc)2, and laminaripentaose β-1-3-(Glc)5 at a concentration of 1.5 mg·mL−1 were used as standards (Megazyme, Bray, Ireland). 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