To solve this problem, a microfluidic circuit has been fabricated in a three-layer glass-PDMS-glass device. In vitro evolution of RNA molecules requires a method for executing many consecutive serial dilutions. The method maintains this accuracy even in the presence of dilution errors of up to 10% (for both the aliquot and diluent volumes), microbial counts between 10(4) and 10(12) colony-forming units, dilution ratios from 2 to 100, and plate size to colony size ratios between 6.25 to 200. The proposed approach shows relative accuracy well within Â☐.1log10 from data produced by computer simulations. The required inputs are the plate size, the microbial colony size, and the serial dilution factors. The estimate of the optimal count given by our method can be used to narrow the search for the best (optimal) dilution plate and saves time. Our method selects the best agar plate with which to estimate the microbial counts, and takes into account the colony size and plate area that both contribute to the likelihood of miscounting the number of colonies on a plate. The number (concentration) of viable microbial organisms is estimated from a single dilution plate (assay) without a need for replicate plates. A simple method is presented to estimate the microbial counts obtained with the serial dilution technique for microorganisms that can grow on bacteriological media and develop into a colony. Titration of microorganisms in infectious or environmental samples is a corner stone of quantitative microbiology. Estimation method for serial dilution experiments.
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