Application of T-RFLP analysis for bacterial community structure of colonies grown on agar plates
2003
Matsuka, A. (Kyushu Univ., Fukuoka (Japan). Faculty of Agriculture) | Sakai, M. | Kanazawa, S.
As a prompt and efficient method for analysis of community structure of bacterial colonies grown on agar plates, we investigated the application of terminal restriction fragment length polymorphism (T-RFLP) analysis. To deduce the community structure of colonies grown on agar plates, analysis of a number of colonies is necessary. In this analytical method, many colonies were collected using a Pasteur pipette to easily collect a constant amount of cells, and the colonies were collectively subjected to T-RFLP analysis to estimate the community structure of colonies. To analyze dominant bacterial species in test soils, 100 colonies were sufficient. Using this analytical method, we investigated the influence of plating method for colony count (pour-plate method vs. spread-plate method) on the constitution of bacterial species grown on the medium. Difference in plating method did not markedly change the bacterial species grown, but it did affect the constitution ratio of some bacterial species.
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