transesterification - MetabolicEngineeringGroupCBMA/MetabolicEngineeringGroupCBMA.github.io GitHub Wiki

Methylation or Trans-esterification (Lund, Dept of Biotechnology)

Prepare 2% Sulfuric acid in methanol (2 mL concentrated H2SO4 in 100 mL Methanol). Add the acid slowly in a volumetric flask as heat is produced. This solution is good for several weeks if kept in fridge.

Add 2 mL of the acid methanol to each tube with dry sample.

Incubate at 80°C for 45 min (heat block)

Remove tube from heat

Add 2 mL MQ water

Invert tubes a couple of times

Transfer to larger glass tubes by decanting

Rinse small tube with 2 mL heptane, add to the large tube by decanting

Vortex large tube for 10s

Centrifuge 1500g for 8 min for phase separation

Transfer upper (organic) phase to a large HPLC vial