title: Protocol: a medium-throughput method for determination of cellulose content from single stem pieces of Arabidopsis thaliana.
abstract: BACKGROUND
  Lignocellulosic biomass is an important renewable resource for biofuels and materials. How plants synthesise cellulose is not completely understood. It is known that cellulose synthase complex (CSCs) moving in the plasma membrane synthesise the cellulose. CESA proteins are the core components of CSC. In Arabidopsis, in vitro mutagenesis of proteins followed by complementation analysis of mutants lacking the gene represents an important tool for studying any biological process, including cellulose biosynthesis. Analysis of a large number of plants is crucial for these types of studies.
  
  
  RESULTS
  By using aspiration rather than centrifugation to remove liquids during various stages of protocol, we were able to increase the throughput of the method as well as minimise the sample loss. As a test case, we determined cellulose content of wild type and secondary wall cesa mutants across the length of primary shoot which was fond to be rather uniform in 7-week-old plants. Additionally, we found that the cellulose content of single mutants was comparable to the higher order mutants.
  
  
  CONCLUSIONS
  Here we describe a medium-throughput adaptation of Updegraff's method that allowed us to determine cellulose content of 200 samples each week.
authors.affiliation: Faculty of Life Sciences, University of Manchester, The Michael Smith Building, Oxford Road, Manchester, M13 9PT UK.
authors.forename: Manoj
authors.initials: M
authors.lastname: Kumar
authors.affiliation: Faculty of Life Sciences, University of Manchester, The Michael Smith Building, Oxford Road, Manchester, M13 9PT UK.
authors.forename: Simon
authors.initials: S
authors.lastname: Turner
keywords: CESA proteins; Cellulose assay; High-throughput; irx mutants