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(3) Analysis of the total peptides synthesizied in the wheat germ assay by sodium dodecyl sulfate polyacrylamide gel electrophoresis demonstrated the presence of a single radioactive peak that corresponded exactly to a specifically immunoprecipitable ovalbumin standard.
This inhibition was specific, since QC did not inhibit the translation of the capped/polyadenylated p33 mRNA in the wheat germ assay (Fig. 4).
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To test this possibility, we used the in vitro wheat germ translation assay that can be programmed with added RNA templates.
For diagnosis, the isolates were first tested by the germ tube assay.
Isolates positive in germ tube assay were checked for growth at 42°C to differentiate C. albicans from C. dubliniensis [ 21].
In this regard, rolA mRNA appears to be poorly translated in a wheat germ translation assay [ 18, 19].
The VITEK Yeast Biochemical Card (YBC, bioMerieux, St . Louis MI, USA) was then used to analyze isolates appearing to be negative by the germ tube assay in the NHRI laboratory and isolates identified as non-albicans Candida species by the hospitals.
In general, isolates identified as C. albicans by hospitals were first subjected to the germ tube assay in brain heart infusion (BHI, BBL) medium containing 10% fetal bovine serum (JR12003, JRH Biosciences, Australia) at 37°C for 2 3 hours [ 20].
To illustrate the step-by-step protocol followed to assay germ cell transplantation into the seminiferous epithelium of mouse testes.
Wheat germ-based thioester assay of UBC1.
Additional file 1: Wheat germ-based thioester assay of UBC1.
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