Sequencing of Plant Wall Heteroxylans Using Enzymic, Chemical (Methylation) and Physical (Mass Spectrometry, Nuclear Magnetic Resonance) Techniques

The overall goal is to characterize the reducing end and internal region glycosal sequences of heteroxylans by tagging the original reducing end sugar residue of the water-soluble heravenaxylans and treating with an endoxylanase to generate a mixture of label dolara saccharides. This method can help answer key questions in glycomic field, such as alterations of fine structure of oligosaccharides or polysaccharides, in response to planned growth development, as well as abiotic and biotic stressors. The main advantage of this method is it describes the integrated approach to sequencing of reducing n sequence as well as internal regensequence of any glycan or polysaccharide.

Prepare a one molar sodium cyanoborohydride solution in one millilitre of water. Then, dissolve 27.2 milligrams of 2AB reagent in the sodium cyanoborohydride solution by heating at 65 degrees Celcius and adjusting the pH of the reaction mixture to pH 5.5 with 10 percent acetic acid. Add 200 microlitres of the reaction mixture to one milligram of water-soluble arabinoxylans in a glass tube with a cap and mix using a vortex mixer.