Efficient Cell Wall Composition Analysis

Recently, there have been methods published that report comprehensive analysis of cell wall components; for example Pettolino et al. [14 (link)] and Foster et al. [15 (link)]. Most of these methods usually involve relatively larger amounts of cell wall material which is ground into a fine powder. For example, Foster et al. [15 (link)] typically start with 60–70 mg dried AIR. Our analysis on the other hand typically stats with 10 mg stem material for wild type samples and 2 mg for irx mutants. The smaller amount of starting sample amounts and the deliberate avoiding of grinding steps to increase throughput means that not all of the side-analyses can be incorporated into our method. However, we do anticipate that an extraction of intact stems with 2 M trifluoroacetic acid (TFA) prior to acetic/nitric treatment can be performed to yield material for analysis of non-cellulosic polysaccharides with GC–MS [15 (link)].
The time of some steps could be further reduced with the use of additional equipment. For example, although our method means it is convenient to carryout the drying steps overnight; a vacuum desiccator or speed vac could be used for the drying steps to reduce time. Another potential time saving improvement could be performing the anthrone assay in a 96-well plate format. This will need an appropriate plate reader to be available which could be used with strong acids (67 % sulphuric acid). Additionally, it will also need 96-well plates that are resistant to 67 % acid at high temperature. However, both these variations have been previously used [15 (link)].