A laboratory experiment was conducted to evaluate the effect of triphenyltetrazolium chloride (TTC)on soil microorganisms and the availability of pH characterization medium in BIOLOG plates. Applicationof TTC decreased the color development sharply and resulted in a great biocidal effect on the growth andreproduction of soil microorganisms, indicating that TTC can affect the discrimination on soil microbialcommunity. The microtitration plates with 21 carbon sources and two different pH levels (4.7 and 7.0) wereused to determine microbial community structure of eight red soils. The average utilization (average wellcolour development) of the carbon sources in the plates with different pH levels generally followed the samesigmoidal pattern as that in the traditional BIOLOG plates, but the pH 4.7 plates increased the discrimination of this technique, compared with the pH 7.0 plates. Since most tested soils are acid, it seemed that its better to use a suitable pH characterization medium for a specific soil in the sole carbon source test.