The long term goal of this project is to understand the symbiosis among gut microbes and the current goal of the project is to develop a quantitative analysis method to measure the growth of specific gut microbes including Escherichia coli HS and Lactobacillus Rhamnosus GG in cell mass or at an RNA level. In this paper, we discussed the early stage of our development including the growth study of the target bacteria using UV-VIS spectrophotometry and quantitative PCR analysis. The specific strains of the two target bacteria in use have been identified using common 16S rRNA sequencing. The growth studies of both bacteria were completed under aerobic and anaerobic conditions. For the UV-VIS spectrophotometry analysis, an OD600 measurement was used and the result clearly showed the differences in the growth phases of the two different bacterial strains under the same growth conditions (Tryptic soy broth media, 37°C and aerobic or anaerobic). For the QPCR analysis, a 16S rRNA template was employed and the results showed similar outcomes with the growth studies using the UV-VIS spectrophotometry for E.coli HS. Currently, we are trying to connect the two different quantitative bacterial growth analysis techniques including cell mass analysis using UV-VIS spectrophotometry and quantification of 16S rRNA using quantitative PCR. In the future, this combination of the two different methods may provide us one of the possible tools to quantitate a specific bacteria strain from a mixture of different microbes using strain specific PCR primers.

Bacterial growth studies of gut microbes including Lactobacillus rhamnosus GG and Escherichia coli HS using UV-VIS spectrophotometry and Quantitative PCR (QPCR)

253rd National Meeting (2017)

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