Table 1 Summary of the changes from the original MBTH1 method to be adapted to the HT2 format
From: Automated saccharification assay for determination of digestibility in plant materials
Reagent | Original method | Modification for HT assay | ||
|---|---|---|---|---|
Volume of reagent 3 | Concentration 3 | Volume of reagent 4 | Concentration 3 | |
Sample | 100 μl | - | 75 μl | - |
NaOH | 100 μl | 0.5 N | 25 μl | 1 N |
MBTH mixture | 100 μl | 1.5 mg/ml MBTH 0.5 mg/ml DTT | 50 μl | 0.21 mg/ml MBTH 0.7 mg/ml DTT |
Oxidising Reagent | 200 μl | 0.5% (FeNH4(SO4)2) 0.5% Sulfamic acid 0.25 N HCl | 100 μl | 0.5% (FeNH4(SO4)2) 0.5% Sulfamic acid 0.25 N HCl |
H2O | 500 μl | - | - | - |
Heating | 80°C for 15 min - | 60°C for 20 min - | ||
Range of detection | 0 to 20 nmol | 0 to 200 nmol | ||